Carbapenem antibacterials with gram-negative activity and processes for their preparation

ABSTRACT

The present invention provides β-methyl carbapenem compounds and pharmaceutical compositions useful in the treatment of bacterial infections and methods for treating such infections using such compounds and/or compositions. The invention includes administering an effective amount of a carbapenem compound or salt and/or prodrug thereof to a host in need of such a treatment. The present invention is also in the field of synthetic organic chemistry and is specifically provides an improved method of synthesis of β-methyl carbapenems which are useful as antibacterial agents.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Application No.60/578,632, filed Jun. 10, 2004.

FIELD OF THE INVENTION

This application provides novel carbapenem compounds and their salts andprodrugs, methods of treatment of gram negative bacterial infectionswith an effective amount of the compounds and pharmaceuticalcompositions including the compounds.

DESCRIPTION OF RELATED ART

The worldwide exploitation of antibiotics to treat infectious diseaseshas grown dramatically over the last forty years. In 1954, two millionpounds of antibiotics were produced in the United States. Today, thefigure exceeds 50 million pounds. According to the Centers DiseaseControl (CDC), humans consume 235 million doses of antibiotics annually.

Widespread misuse or overuse of antibiotics has fostered the spread ofantibiotic resistance and has contributed to the development of aserious public health problem. Antibiotic resistance occurs whenbacteria that cause infection are not killed by the antibiotics taken tostop the infection. The bacteria survive and continue to multiply,causing more harm. For example, the bacterium Staphylococcus aureus is amajor cause of hospital acquired infections that, historically,responded satisfactorily to the antibiotic vancomycin. Recently,however, many strains of S. aureus have been found to be resistant tovancomycin. Moreover, the death rate for some communicable diseases suchas tuberculosis have started to rise again, in part because of increasesin bacterial resistance to antibiotics.

Antibiotics are used therapeutically to treat bacterial infections.Several types of antibiotics, classified according to their mechanism ofaction, are currently employed. The known types of antibiotics include,e.g. cell wall synthesis inhibitors, cell membrane inhibitors, proteinsynthesis inhibitors and inhibitors that bind to or affect the synthesisof DNA or RNA.

Cell wall synthesis inhibitors, such as beta lactam antibiotics,generally inhibit some step in the synthesis of bacterial peptidoglycan.Penicillin is generally effective against non-resistant streptococcus,gonococcus and staphylococcus. Amoxycillin and Ampicillin have broadenedspectra against Gram-negative bacteria. Cephalosporins are generallyused as penicillin substitutes, against Gram-negative bacteria and insurgical prophylaxis. Monobactams are generally useful for the treatmentof allergic individuals.

Numerous antibiotic agents, suitable for use in the treatment ofbacteria-related diseases and disorders, are known and disclosed, e.g.in The Physician's Desk Reference (PDR), Medical Economics Company(Montvale, N.J.), (53^(rd) Ed.), 1999; Mayo Medical Center Formulary,Unabridged Version, Mayo Clinic (Rochester, Minn.), January 1998; MerckIndex: An Encyclopedia of Chemicals, Drugs and Biologicals, (11^(th)Ed.), Merck & Co., Inc. (Rahway, N.J.), 1989; University of WisconsinAntimicrobial Use Guide,http://www.medsch.wisc.edu/clinsci/5amcg/amcg.html; Introduction on theUse of the Antibiotics Guideline, of Specific Antibiotic Classes, ThomasJefferson University,http://jeffiine.tju.edu/CWIS/OAC/antibiotics_guide/intro.html; andreferences cited therein.

The first carbapenem to be isolated was thienamycin, shown below, whichwas isolated from Streptomyces cattleya (U.S. Pat. No. 3,950,357) andwas shown to have strong antibacterial activity, including potencyagainst Pseudomonas spp. and β-lactamase stability (Kahan, J. S., etal., J. Antibiot., 32, pp. 1-12 (1979); Bodey, G. P., et al.,Antimicrob. Agents Chemother., 15, pp. 518-521 (1979). The racemicsynthesis of thienamycin was reported shortly thereafter by Merck(Johnston, D. B. R., et al., J. Am. Chem. Soc., 100, pp. 313-315 (1978);Bouffard, F. A., et al., J. Org. Chem., 45, 1130-1142 (1980)), as wellas an asymmetric total synthesis (Salzmann, T. N., et al., J. Am. Chem.Soc. 102, pp. 6161-6163 (1980)). The nucleus and amino-containing sidechain of this molecule,

however, contributed to its chemical instability. In addition to itspotential to be hydrolyzed by the zinc-activated β-lactamase that ispresent in Bacillus species, Xanthomonas, Pseudomonas, and Bacteroidesspecies (Saino, Y., et al., Antimicrob. Agents Chemother., 22, pp.564-570 (1982); Yotsujii, A., et al., Antimicrob. Agents Chemother., 24,pp. 925-929 (1983)), chemical stability issues associated with theintermolecular aminolysis of the azetidinone (β-lactam) ring of onemolecule of thienamycin by the primary amine in the cysteamine sidechain of another thienamycin molecule, resulted in the use ofthienamycin as a drug candidate to be abandoned.

As a result of the problems associated with thienamycin, N-formimidoylthienamycin, known as imipenem, was synthesized (Leanza, W. J., et al.,J. Med. Chem., 22, pp. 1435-1436 (1979)). This compound bears a morebasic amidine functionality on the 2′ side chain, which is protonated atphysiological pH, preventing the compound from initiating a nucleophilicattack on another imipenem molecule.

However, poor urinary tract recovery from test subjects revealed aninstability of this compound to the mammalian β-lactamase renaldehydropeptidase-I (DHP-I) (Shimada, J., et al., Drugs Exp Clin Res.,20, pp. 241-245 (1994)). Consequently, the compound cilastatin wasdeveloped for use in co-administration in order to prevent hydrolysisand degredation by DHP-I; this combination therapy is currentlyprescribed under the name Primaxin® (Merck Frosst Std).

In response to the problem of carbapenems to destruction by renaldehydropeptidase-1, the carbapenem antibiotic meropenem (SM7338) (shownbelow), was developed (see, Edwards, J. R., et al., Antimicrob. AgentsChemother., 33, pp. 215-222 (1989); Neu, H. C., et al., Antimicrob.Agents Chemother., 33, pp. 1009-1018 (1989)).

This compound was shown to be active against a large number ofGram-negative bacteria. The drug is currently prescribed for intravenoususe (Merrem® IV; AstraZeneca) in the treatment of intra-abdominalinfections and bacterial meningitis.

The carbapenem ertapenem (formerly MK-0826; Cunha, B. A., Drugs ofToday, 38, pp. 195-213 (2002)) was the first of a group of carbapenemswith potential against methicillin-resistant staphylococci (MRS) shownto be useful as a long-acting, parenteral carbapenem (Shah, P. M., etal., J. Antimicrob. Chemother., 52, pp. 538-542 (2003); Aldridge, K. E.,Diagn. Microbiol. Infect. Dis., 44(2), pp. 181-6 (2002)). It is suitablefor administration both as a single-agent (e.g., co-administration witha compound such as cilastatin is not required), or by the intravenous orintramuscular route (Legua, P., et al., Clin. Therapeut., 24, pp.434-444 (2002); Majumdar, A. K., et al., Antimicrob. Agents Chemother.,46, pp. 3506-3511 (2002)). Eratapenem has received regulatory approvalin both the United States (November, 2001) and the European Union(April, 2002).

One carbapenem having a fused pyrazole ring system (L-627; Biapenem) wasdeveloped by Lederle Ltd. (Japan), and introduced a methyl radical atthe 1-β position of the carbapenem skeleton (see, U.S. Pat. No.4,866,171). This structural modification reportedly gave biapenemstability against hydrolysis by kidney dehydropeptidase, makingco-administration of a dehydropeptidase inhibitor unnecessary.

More recently, a new, injectable 1-β-methyl carbapenem antibiotic havingan (R)-1-hydroxymethyl-methylaminopropyl group exhibiting both broadspectrum, potent antibacterial activity (BO-2727) and havingantipseudomonal activity has been reported (Nakagawa, S., et al.,Antimicrob. Agents Chemother., 37, pp. 2756-2759 (1993); Hazumi, N., etal., Antimicrob. Agents Chemother., 39, pp. 702-706 (1995

Since the discovery of thienamycin having a potential antimicrobialactivity against Gram-negative and Gram-positive bacteria, studies onthe syntheses of carbapenem derivatives which are analogous tothienamycin have been widely developed. As a result, it was found thatcarbapenem derivatives having, as their 2-side chain, a substituentderived from 4-hydroxy-proline exhibit a potential antimicrobialactivity and are useful as medicines or as intermediates for compoundspossessing antimicrobial activity.

1-β-methyl carbapenem antibiotics, are particularly well known fortreating a broad spectrum of gram-negative and gram-positive bacterialinfections. See for example U.S. Pat. No. 4,962,103; U.S. Pat. No.4,933,333; U.S. Pat. No. 4,943,569; U.S. Pat. No. 5,122,604; U.S. Pat.No. 5,034,384 and U.S. Pat. No. 5,011,832.

U.S. Pat. No. 6,255,300 to Merck & Co. describes certain carbapenemantibacterial agents in which the carbapenem nucleus is substituted withan iodo-phenyl linked through a methyl-oxygen lineage. The patent statesthat these compounds are useful against gram positive bacterialinfections. Similarly, U.S. Pat. No. 6,310,055 provides carbapenemcompounds with aromatic side chains that are halogen substituted, linkedthrough an alkoxy unsaturated group.

European Publication No. 0 292 191 to Merck & Co. describes certain2-(substituted methyl)-1-alkylcarbapenem compounds useful as antibioticagents.

U.S. Pat. No. 6,399,597, also to Merck & Co. describes certainnaphthosultam compounds that are allegedly useful in the treatment ofcertain drug resistant bacterial infections.

Because of the difficulty in developing effective carbapenem compoundsdue to hydrolysis of the β-lactam ring and low recovery, compounds withsuperior anti-bacterial activity have not been developed.

Therefore, it is one object of the present invention to provide novelβ-methyl compounds carbapenems that are effective antimicrobial agents.

It is another object of the present invention to provide methods for thetreatment of gram-negative bacteria, that is optionally can bedrug-resistant and/or multi-drug resistant.

SUMMARY OF THE INVENTION

In one embodiment of the present invention, carbapenems of the generalformula (I)

or a pharmaceutically acceptable salt and/or prodrug thereof aredescribed, wherein

-   R is H or alkyl, typically lower alkyl such as CH₃;-   R¹ is H or alkyl, typically lower alkyl such as CH₃;-   M is H or a group such that CO₂M represents a carboxylic acid, a    carboxylate anion, a pharmaceutically acceptable ester group, or a    carboxylic acid protected by a protecting group;-   P is selected from the group consisting of hydrogen, hydroxyl,    halogen (such as F), or hydroxyl protected by a hydroxyl protecting    group; and-   each Y¹ and Y² is independently selected from the group consisting    of hydrogen; halo; —CN; —NO₂; —NR^(a)R^(b); —OR^(c); —SR^(c);    —C(O)NR^(a)R^(b); —C(O)OR^(h); S(O)R^(c); —SO₂R^(c);    —SO₂NR^(a)R^(b); —NR^(a)SO₂R^(b); —C(O)R^(a); —OC(O)R^(a);    OC(O)NR^(a)R^(b); —NRaC(O)NR^(b)R^(c); —NR^(a)CO₂R^(h); —OCO₂R^(h);    NR^(a)C(O)R^(b); C₁₋₆ straight- or branched-chain alkyl that are    substituted or unsubstituted with one to four R^(d) groups; C₁₋₆    straight- or branched-chain alkyl that are substituted or    unsubstituted one to four R^(d) groups and with or without    saturation (double or triple bonds); -A-(CH₂)_(n)-Q and C₃₋₇    cycloalkyl, substituted or unsubstituted with one to four R^(d)    groups;-   wherein A is selected from the group consisting of O, S, NH, NCH₃,    NR, or —CH₂—;-   n is 0, 1, 2 or 3;-   each R^(a), R^(b) and R^(c) is independently selected from hydrogen,    —C₁₋₆ straight- or branched-chain alkyl, unsubstituted or    substituted with one to four R^(d) groups, or —C₃₋₇ cycloalkyl,    unsubstituted or substituted with one to four R^(d) groups;-   or R^(a) and R^(b) taken together with any intervening atoms    represent a 4-6 membered saturated ring optionally interrupted by    one or more of O, S, NR^(c), with R^(c) as defined above, or —C(O)—,    said ring being unsubstituted or substituted with one to four R^(i)    groups;-   or R^(b) and R^(c) taken together with any intervening atoms    represent a 4-6 membered saturated ring optionally interrupted by    one to three of O, S, NR^(a), with R^(a) as defined above, or    —C(O)—, said ring being unsubstituted or substituted with one to    four R^(i) groups;-   each R^(d) is independently selected from the group consisting of    halogen; —CN; —NO₂; —NR^(e)R^(f); —OR^(g); —SR^(g); —CONR^(e)R^(f);    —COOR^(g); —SOR^(g); —SO₂R^(g); —SO₂NR^(e)R^(f); —NR^(e)SO₂R^(f);    —COR^(e); —NR^(e)COR^(f); —OCOR^(e); —OCONR^(e)R^(f)l    NR^(e)CONR^(f)R^(g); —NR^(e)CO₂R^(h); —OCO₂R^(h);    —C(NR^(e))NR^(f)R^(g); NR^(e)C(NH)NR^(f)R^(g) or    —NR^(e)C(NR^(f))R^(g);-   each R^(e), R^(f) and R^(g) independently represents hydrogen; —R;    —C₁₋₆ straight- or branched-chain alkyl unsubstituted or substituted    with one to four R^(i) groups;-   or R^(e) and R^(f) taken together with any intervening atoms    represent a 4-6 membered saturated ring optionally interrupted by    one to three of O, S, —C(O)— or NR^(g) with R^(g) as defined above,    said ring being unsubstituted or substituted with one to four R^(i)    groups;-   each R^(i) independently represents halo; —CN; —NO₂; phenyl;    —NHSO₂R^(h); —OR^(h), —SR^(h); —N(R^(h))₂; —N⁺(R^(h))3;    —C(O)N(R^(h))₂; SO₂N(R^(h))₂; heteroaryl; heteroarylium; —CO₂R^(h);    —C(O)R^(h); —OCOR^(h); NHCOR^(h); guanidinyl; carbamimidoyl or    ureido;-   each R^(h) independently represents hydrogen, a —C₁₋₆ straight or    branched-chain alkyl group, a —C₃-C₆ cycloalkyl group or phenyl, or    when two R^(h) groups are present, said R^(h) groups may be taken in    combination and represent a 4-6 membered saturated ring, optionally    interrupted by one or two of O, S, SO₂, —C(O)—, NH and NCH₃;-   each Q is selected from the group consisting of:

wherein.

-   -   a and b are 1, 2 or 3;    -   L⁻ is a pharmaceutically acceptable counterion;    -   α is selected from O, S or NR^(s);    -   each β, δ, λ, μ, and σ is independently selected from CR^(t), N        or N⁺R^(s), provided that no more than one of β, δ, λ, μ, and σ        is N⁺R^(s);    -   each R^(s) is independently selected from hydrogen; phenyl or        C₁₋₆ straight- or branched-chain alkyl, unsubstituted or        substituted with one to four R^(i) groups;    -   each R^(t) is independently selected from hydrogen; halo;        phenyl; —CN; —NO₂; —NR^(u)R^(v); —OR^(u); —SR^(u);        —CONR^(u)R^(v); —COOR^(h); —SOR^(u); —SO₂R^(u); —SO₂NR^(u)R^(v);        —NR^(u)SO₂R^(v); —COR^(u); —NR^(u)COR^(v); —OCOR^(u);        —OCONR^(u)R^(v); —NR^(u)CO₂R^(v); —NR^(u)CONR^(v)R^(w);        —OCO₂R^(v); —C₁₋₆ straight- or branched-chain alkyl,        unsubstituted or substituted with one to four R^(i) groups;    -   each R^(u) and R^(v) is independently hydrogen or —C₁₋₆        straight- or branched-chain alkyl, unsubstituted or substituted        with one to four R^(i) groups;    -   or R^(u) and R^(v) together with any intervening atoms represent        a 4-6 membered saturated ring optionally interrupted by one or        more of O, S, NRW or —C(O)—, said ring being unsubstituted or        substituted with one to four R^(i) groups;    -   each R^(w) independently represents hydrogen; —C₁₋₆ straight- or        branched-chain alkyl, unsubstituted or substituted with one to        four R^(i) groups; C₃₋₆ cycloalkyl optionally substituted with        one to four R^(i) groups; phenyl optionally substituted with one        to four R^(i) groups, or heteroaryl optionally substituted with        1-4 R^(i) groups; or R^(h) and R^(w) taken together with any        intervening atoms represent a 5-6 membered saturated ring,        optionally interrupted by one or two of O, S, SO₂, NH or NCH₃;    -   each R^(x) independently represents hydrogen or a C₁₋₈ straight-        or branched chain alkyl, optionally interrupted by one or two of        O, S, SO, SO₂, NR^(w), N⁺R^(h)R^(w), or —C(O)—, said chain being        unsubstituted or substituted with one to four of halo, CN, NO₂,        OR^(w), SR^(w), SOR^(w), SO₂R^(w), NR^(h)R^(w), N⁺(R^(h))₂R^(w),        —C(O)—R^(w), C(O)NR^(h)R^(w), SO₂NR^(h)R^(w), CO₂R^(w),        OC(O)R^(w), OC(O)NR^(h)R^(w), NR^(h)C(O)R^(w),        NR^(h)C(O)NR^(h)R^(w), or a phenyl or heteroaryl group which is        in turn optionally substituted with from one to four R^(i)        groups or with one to two C₁₋₃ straight- or branched-chain alkyl        groups, said alkyl groups being unsubstituted or substituted        with one to four R^(i) groups;    -   each R^(y) and R^(z) is independently selected from hydrogen;        phenyl; —C₁₋₆ straight or branched chain alkyl, unsubstituted or        substituted with one to four R^(i) groups, and optionally        interrupted by O, S, NR^(w), N⁺R^(h)R^(w) or —C(O)—;    -   or R^(x) and R^(y) together with any intervening atoms represent        a 4-6 membered saturated ring optionally interrupted by O, S,        SO₂, NR^(w), N⁺R^(h)R^(w) or —C(O)—, unsubstituted or        substituted with 1-4 R^(i) groups,    -   and when R^(x) and R^(y) together represent a 4-6 membered ring        as defined above, R^(z) is as defined above or R^(z) represents        an additional saturated 4-6 membered ring fused to the ring        represented by R^(x) and R^(y) taken together, optionally        interrupted by O, S, NR^(w) or —C(O)—, said rings being        unsubstituted or substituted with one to four R^(i) groups.

The present invention also provides a pharmaceutical compositionincluding a compound of the invention, or a pharmaceutically acceptablesalt and/or prodrug thereof, optionally with a pharmaceuticallyacceptable carrier or diluent.

In one embodiment, the invention provides a pharmaceutical compositioncomprising a compound of the invention, or a pharmaceutically acceptablesalt and/or prodrug therein, in combination with one or more otherantimicrobial agents, optionally with a pharmaceutically acceptablecarrier or diluent.

In another embodiment, the invention provides a method of preventing ortreating a bacterial infection in a host, typically an animal, and mosttypically a human, including administering to the host a therapeuticamount of a compound of the present invention, or a pharmaceuticallyacceptable salt and/or prodrug therein, optionally in a pharmaceuticallyacceptable carrier or diluent.

In a separate embodiment, the invention provides a method of preventingor treating a gram-negative bacterial infection in a host that includesadministering a therapeutic amount of a compound of the presentinvention, or a pharmaceutically acceptable salt and/or prodrug therein,in combination or alternation with one or more other antimicrobialagents, optionally in a pharmaceutically acceptable carrier or diluent.

In one principal embodiment, the bacterial infection is due to agram-negative bacteria. In another embodiment, the bacterial infectionis from a drug resistant and/or multiple-drug resistant gram-negativebacteria.

The invention also provides a compound of the present invention for usein medical therapy, and the use in the preparation of a medicament forthe treatment of bacterial infections, particularly gram negativebacterial infections, alone or in combination with another agent.

DESCRIPTION OF THE FIGURES

FIG. 1 shows a nonlimiting illustrative example of known carbapenems.

FIG. 2 shows a nonlimiting illustrative example of the structure ofcarbapenem analogs of the present invention possessing gram-negativebiological activity.

FIG. 3 shows the synthetic process of preparing carbapenem Intermediate5.

FIG. 4 is a table showing gram-negative MIC (In Vitro Susceptibility)Data for selected compounds against selected organism.

DETAILED DESCRIPTION OF THE INVENTION

The invention provides carbapenem compounds or their pharmaceuticallyacceptable salts or prodrugs, pharmaceutical compositions containingthese compounds and methods of their use in the treatment or preventionof gram-negative bacterial infections.

DEFINITIONS

The numbering system for the carbapenem compounds used in thisspecification is set out below, wherein the numbering of the carbapenemnucleus is in accordance with standards in the art (see, Tiraby, G., etal., Biochem J, 276 (pt. 1), pp. 269-270 (1991)).

Whenever a range is presented herein it should be understood to includeeach element of the range. For example, the range “C₁ to C₄” alkylindependently includes C₁, C₂, C₃ and C₄ alkyl groups. When such a rangeis stated, each element has been contemplated and the range is usedmerely for convenience.

Generally, while the compounds, compositions and methods are describedin terms of “comprising” various components or steps, the compounds,compositions and methods can also “consist essentially of” or “consistof” the various components and steps.

The term “alkyl”, as used herein, unless otherwise specified, includes asaturated straight, branched, or cyclic, primary, secondary, or tertiaryhydrocarbon of C₁ to C₁₀. The term includes both substituted andunsubstituted alkyl groups. Moieties with which the alkyl group can besubstituted are selected from the group consisting of hydroxyl, halo (F,Cl, Br, I), amino, alkylamino, arylamino, alkoxy, aryloxy, nitro, cyano,sulfonic acid, sulfate, phosphonic acid, phosphate, or phosphonate,either unprotected, or protected as necessary, as known to those skilledin the art, for example, as taught in Greene, et al., Protective Groupsin Organic Synthesis, John Wiley and Sons, Second Edition, 1991, herebyincorporated by reference. When the alkyl group is said to besubstituted with an alkyl group, this is used interchangeably with“branched alkyl group”. Specific examples of alkyls and/or substitutedalkyls includes, but are not limited to, methyl, trifluoromethyl, ethyl,propyl, isopropyl, cyclopropyl, butyl, isobutyl, t-butyl, pentyl,cyclopentyl, isopentyl, neopentyl, hexyl, isohexyl, cyclohexyl,cyclohexylmethyl, 3-methylpentyl, 2,2-dimethylbutyl, and2,3-dimethylbutyl.

The term “lower alkyl”, as used herein, and unless otherwise specified,refers to a C₁ to C₄ saturated straight, branched, or if appropriate, acyclic (for example, cyclopropyl) alkyl group, including bothsubstituted and unsubstituted forms. Unless otherwise specificallystated in this application, when alkyl is a suitable moiety, lower alkylis typical. Similarly, when alkyl or lower alkyl is a suitable moiety,unsubstituted alkyl or lower alkyl is typical.

Cycloalkyl is a species of alkyl containing from 3 to 15 carbon atoms,without alternating or resonating double bonds between carbon atoms. Itmay contain from 1 to 4 rings which are fused.

The term “alkenyl” includes a hydrocarbon radical straight, branched orcyclic containing from 2 to 10 carbon atoms and at least one carbon tocarbon double bond. Examples of alkenyl groups include ethenyl,propenyl, butenyl and cyclohexenyl.

The term “alkynyl” refers to a hydrocarbon radical straight or branched,containing from 2 to 10 carbon atoms and at least one carbon to carbontriple bond. Examples of alkynyl groups include ethynyl, propynyl andbutynyl.

“Alkoxy” includes C₁-C₄ alkyl-O—, with the alkyl group optionallysubstituted as described herein.

The term “alkylamino” or “arylamino” refers to an amino group that hasone or two alkyl or aryl substituents, respectively.

“Aryl” refers to aromatic rings e.g., phenyl, substituted phenyl,biphenyl, and the like, as well as rings which are fused, e.g.,naphthyl, phenanthrenyl and the like. An aryl group thus contains atleast one ring having at least 6 atoms, with up to five such rings beingpresent, containing up to 22 atoms therein, with alternating(resonating) double bonds between adjacent carbon atoms or suitableheteroatoms. The typical aryl groups are phenyl, naphthyl andphenanthrenyl. The term includes both substituted and unsubstitutedmoieties. The aryl group can be substituted with one or more moietiesselected from the group consisting of bromo, chloro, fluoro, iodo,hydroxyl, amino, alkylamino, arylamino, alkoxy, aryloxy, nitro, cyano,sulfonic acid, sulfate, phosphonic acid, phosphate, or phosphonate,either unprotected, or protected as necessary, as known to those skilledin the art, for example, as taught in Greene, et al., Protective Groupsin Organic Synthesis, John Wiley and Sons, Second Edition, 1991. Typicalsubstituted aryls include phenyl and naphthyl.

The term “alkaryl” or “alkylaryl” refers to an alkyl group with an arylsubstituent. The term “aralkyl” or “arylalkyl” refers to an aryl groupwith an alkyl substituent.

The term “heteroaryl” or “heteroaromatic”, as used herein, refers to anaromatic group that includes at least one sulfur, oxygen, nitrogen orphosphorus in the aromatic ring. Heteroaryl or heteroaromatic compoundsinclude monocyclic aromatic hydrocarbon group having 5 or 6 ring atoms,or a bicyclic aromatic group having 8 to 10 atoms, containing at leastone heteroatom, O, S or N, in which a carbon or nitrogen atom is thepoint of attachment, and in which one, two or three additional carbonatoms are optionally replaced by a heteroatom selected from oxygen,sulfur or nitrogen heteroatom. Examples of this type are pyrrole,pyridine, oxazole, thiazole and oxazine. Additional nitrogen atoms maybe present together with the first nitrogen and oxygen or sulfur,giving, e.g., thiadiazole. Examples include the following.

The heteroaryl or heteroaromatic group can be optionally substitutedwith one or more substituent selected from halogen, haloalkyl, alkyl,alkoxy, hydroxy, carboxyl derivatives, amido, amino, alkylamino,dialkylamino. Functional oxygen and nitrogen groups on the heterocyclicor heteroaryl group can be protected as necessary or desired. Suitableprotecting groups are well known to those skilled in the art, andinclude trimethylsilyl, dimethylhexylsilyl, t-butyldimethylsilyl, andt-butyl-diphenylsilyl, trityl or substituted trityl, alkyl groups, acylgroups such as acetyl and propionyl, methanesulfonyl, andp-toluenylsulfonyl.

“Heteroarylium” refers to heteroaryl groups bearing a quaternarynitrogen atom and thus a positive charge. Examples include thefollowing.

When a charge is shown on a particular nitrogen atom in a ring whichcontains one or more additional nitrogen atoms, it is understood thatthe charge may reside on a different nitrogen atom in the ring by virtueof charge resonance that occurs.

The term “heterocycloalkyl” refers to a cycloalkyl group (nonaromatic)in which one of the carbon atoms in the ring is replaced by a heteroatomselected from O, S or N, and in which up to three additional carbonatoms may be replaced by heteroatoms.

The terms “quaternary nitrogen” and “positive charge” refer totetravalent, positively charged nitrogen atoms including, e.g., thepositively charged nitrogen in a tetraalkylammonium group (e.g.tetramethylammonium), heteroarylium, (e.g., N-methyl-pyridinium), basicnitrogens which are protonated at physiological pH, and the like.Cationic groups thus encompass positively charged nitrogen-containinggroups, as well as basic nitrogens which are protonated at physiologicpH.

The term “heteroatom” refers to oxygen, sulfur, nitrogen, phosphorus,and selenium, selected on an independent basis.

Halogen and “halo”, as used herein, includes bromine, chlorine, fluorineand iodine.

The term acyl refers to a carboxylic acid ester in which thenon-carbonyl moiety of the ester group is selected from straight,branched, or cyclic alkyl or lower alkyl, alkoxyalkyl includingmethoxymethyl, aralkyl including benzyl, aryloxyalkyl such asphenoxymethyl, aryl including phenyl optionally substituted withhalogen, C₁ to C₄ alkyl or C₁ to C₄ alkoxy, sulfonate esters such asalkyl or aralkyl sulphonyl including methanesulfonyl, the mono, di ortriphosphate ester, trityl or monomethoxytrityl, substituted benzyl,trialkylsilyl (e.g. dimethyl-t-butylsilyl) or diphenylmethylsilyl. Arylgroups in the esters typically include a phenyl group. The term “loweracyl” refers to an acyl group in which the non-carbonyl moiety is loweralkyl.

“Carboxylate anion” refers to a negatively charged group —COO.

“Guanidinyl” refers to the group: H₂NC(NH)NH—.

“Carbamimidoyl” refers to the group: H₂NC(NH)—.

“Ureido” refers to the group: H₂NC(O)NH—.

When a group is “optionally interrupted”, this includes one or more ofthe interrupting moieties in combination, as well as said moietieslocated at either or both ends of the chain. Thus, it includesterminating the group as well.

When a group is termed “substituted”, unless otherwise indicated, thismeans that the group contains from 1 to 4 substituents thereon. Withrespect to R, R^(a), R^(b) and R^(c), the substituents available onalkyl groups are selected from the values of R^(d). Many of the variablegroups are optionally substituted with up to four R^(i) groups. Withrespect to R^(e), R^(f) and R^(g), when these variables representsubstituted alkyl, the substituents available thereon are selected fromthe values of R^(i).

When a functional group is termed “protected”, this means that the groupis in modified form to preclude undesired side reactions at theprotected site, and unless otherwise defined refers to a group that isadded to an oxygen, nitrogen, or phosphorus atom to prevent its furtherreaction or for other purposes. In some of the carbapenem compounds ofthe present invention, M is a readily removable carboxyl protectinggroup, and/or P represents a hydroxyl which is protected by a hydroxylprotecting group. Such protecting groups are used to protectively blockthe hydroxyl or carboxyl group during the synthesis procedures and arereadily removable by procedures that will not cause cleavage or otherdisruption of the remaining portions of the molecule. Such proceduresinclude chemical and enzymatic hydrolysis, treatment with chemicalreducing or oxidizing agents under mild conditions, treatment with atransition metal catalyst and a nucleophile and catalytic hydrogenation.

A wide variety of oxygen and nitrogen protecting groups are known tothose skilled in the art of organic synthesis. Suitable protectinggroups for the compounds of the present invention will be recognizedfrom the present application taking into account the level of skill inthe art, and with reference to standard textbooks, such as Greene, T. W.and Wuts, P. M., Protective Groups in Organic Synthesis, 3^(rd) Ed.,Wiley, New York (1991). Examples of carboxyl protecting groups includeallyl, benzhydryl, 2-naphthylmethyl, benzyl (Bn), silyl such ast-butyldimethylsilyl (TBDMS), phenacyl, p-methoxybenzyl, o-nitrobenzyl,p-methoxyphenyl, p-nitrobenzyl, 4-pyridylmethyl and t-butyl. Examples ofsuitable C-6 hydroxyethyl protecting groups include triethylsilyl (TES),t-butyldimethylsilyl (TBDMS), o-nitrobenzyloxycarbonyl (ONB),p-nitrobenzyloxycarbonyl (PNB), benzyloxycarbonyl (CBz),allyloxycarbonyl (Alloc), t-butyloxycarbonyl (Boc),2,2,2-trichloroethyloxycarbonyl (Troc), and the like.

The phrase “pharmaceutically acceptable ester, salt or hydrate,” refersto those salts, esters and hydrated forms of the compounds of thepresent invention which would be apparent to the pharmaceutical chemist.i.e., those which are substantially non-toxic and which may favorablyaffect the pharmacokinetic properties of said compounds, such aspalatability, absorption, distribution, metabolism and excretion. Otherfactors that are also important in the selection are cost of the rawmaterials, ease of crystallization, yield, stability, solubility,hygroscopicity and flowability of the resulting bulk drug.

“Pharmaceutically acceptable salts” include salts that retain thedesired biological activity of the parent compound and do not impartundesired toxicological effects. These salts can take the form —COOM,where M is a negative charge, which is balanced by a counterion. Theseinclude salts formed with cations such as sodium, potassium, NH₄ ⁺,magnesium, zinc, ammonium, or alkylammonium cations such astetramethylammonium, tetrabutylammonium, choline, triethylhydroammonium,meglumine, triethanolhydroammonium, calcium, and calcium polyamines suchas spermine and spermidine. These can also include salts formed fromelemental anions such as chloride, bromide, and iodide. They can alsoinclude acid addition salts, for example, salts derived from inorganicor organic acids. Included among such salts are the following: acetate,adipate, alginate, ascorbic acid, aspartate, benzoate, benzenesulfonate,bisulfate, butyrate, citrate, camphorate, camphorsulfonate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,fumarate, glucoheptanoate, gluconic acid, glycerophosphate, hemisulfate,heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide,2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitric acid, oxalate, palmitic acid,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphoric acid,picrate, pivalate, polygalacturonic acid; polyglutamic acid, propionate,p-toluenesulfonic acid, succinate, sulfuric acid, tannic acid, tartrate,thiocyanate, tosylate and undecanoate.

The term “prodrug” includes a compound that, when administered to ananimal, is converted under physiological conditions to a compound of theinvention, for example a pharmaceutically acceptable ester.

The pharmaceutically acceptable esters are such as would be readilyapparent to a medicinal chemist, and include, for example, thosedescribed in detail in U.S. Pat. No. 4,309,438. Included within suchpharmaceutically acceptable esters are those which are hydrolyzed underphysiological conditions, such as pivaloyloxymethyl, acetoxymethyl,phthalidyl, indanyl and methoxymethyl. These are also referred to as“biolabile esters”, which are biologically hydrolysable. Examples ofbiolabile esters include compounds in which M represents an alkoxyalkyl,alkylcarbonyloxyalkyl, alkoxycarbonyloxyalkyl, cycloalkoxyalkyl,alkenyloxyalkyl, aryloxyalkyl, alkoxyaryl, alkylthioalkyl,cycloalkylthioalkyl, alkenylthioalkyl, arylthioalkyl or alkylthioarylgroup. These groups can be substituted in the alkyl or aryl portionsthereof with acyl or halo groups. The following M species are examplesof biolabile ester forming moieties: acetoxymethyl, 1-acetoxyethyl,1-acetoxypropyl, pivaloyloxymethyl, lisopropyloxycarbonyloxyethyl,1-cyclohexyloxycarbonyloxyethyl, phthalidyl and (2-oxomethyl-1,3-dioxolenyl)methyl.

The term “host”, as used herein, refers to a unicellular ormulticellular organism in which the bacteria can replicate, includingcell lines and animals. Alternatively, the host can be carrying a partof the bacterial particles, whose replication and/or function can bealtered by the compounds of the present invention. The term host refersto infected cells, cells transfected with all or part of the bacteriaand animals, such as, primates (including chimpanzees) and, in oneembodiment, the host is a human. Veterinary applications are alsoencompassed by the present invention.

The term “treatment” as used herein, includes an approach for obtainingbeneficial or desired results including clinical results, includingalleviation of symptoms, diminishment of extent of disease,stabilization (i.e., not worsening) state of disease, preventing spreadof disease, preventing or reducing occurrence or recurrence of disease,delay or slowing of disease progression, and reduction of incidence ofdisease or symptoms. As used herein, the phrase “anti-bacteriallyeffective amount” means an amount effective for treating the bacterialinfection.

Compounds of the Invention

In one embodiment of the present invention, the carbapenem of theformula I,

or pharmaceutically acceptable salts and/or prodrugs thereof isprovided, whereinR is H or alkyl, such as lower alkyl such as CH₃;R¹ is H or alkyl, such as lower alkyl such as CH₃;M is H or a group such that CO₂M represents a carboxylic acid, acarboxylate anion, a pharmaceutically acceptable ester group, or acarboxylic acid protected by a protecting group;P is hydrogen, hydroxyl, halogen such as F, or hydroxyl protected by ahydroxyl protecting group; andY¹ and Y² are each independently selected from the following: hydrogen;halo; —CN; —NO₂; —NR^(a)R^(b); —OR^(c); —SR^(c); —C(O)NR^(a)R^(b);—C(O)OR^(h); S(O)R^(c); —SO₂R^(c); —SO₂NR^(a)R^(b); —NR^(a)SO₂R^(b);—C(O)R^(a); —OC(O)R^(a); OC(O)NR^(a)R^(b); —NRaC(O)NR^(b)R^(c);—NR^(a)CO₂R^(h); —OCO₂R^(h); NR^(a)C(O)R^(b); C₁₋₆ straight- orbranched-chain alkyl that are substituted or unsubstituted with one tofour R^(d) groups and with or without saturation (double or triplebonds); alkylaryl, -A-(CH₂)_(n)-Q and C₃₋₇ cycloalkyl, substituted orunsubstituted with one to four R^(d) groups.wherein A represents O, S, NH, NCH₃, NR, or —CH₂—;n represents an integer 0, 1, 2 or 3;

-   each R^(a), R^(b) and R^(c) independently represents hydrogen, —C₁₋₆    straight- or branched-chain alkyl, unsubstituted or substituted with    one to four R^(d) groups, or —C₃₋₇ cycloalkyl, unsubstituted or    substituted with one to four R^(d) groups;-   or R^(a) and R^(b) taken together with any intervening atoms    represent a 4-6 membered saturated ring optionally interrupted by    one or more of O, S, NR^(c), with R^(c) as defined above, or —C(O)—,    said ring being unsubstituted or substituted with one to four R^(i)    groups;-   or R^(b) and R^(c) taken together with any intervening atoms    represent a 4-6 membered saturated ring optionally interrupted by    one to three of O, S, NR^(a), with R^(a) as defined above, or    —C(O)—, said ring being unsubstituted or substituted with one to    four R^(i) groups;-   each R^(d) independently represents halo; —CN; —NO₂; —NR^(e)R^(f);    —OR^(g); —SR^(g); —CONR^(e)R^(f); —COOR^(g); —SOR^(g); —SO₂R^(g);    —SO₂NR^(e)R^(f); —NR^(e)SO₂R^(f); —COR^(e); —NR^(e)COR^(f);    —OCOR^(e); —OCONR^(e)R^(f)l NR^(e)CONR^(f)R^(g); —NR^(e)CO₂R^(h);    —OCO₂R^(h); —C(NR^(e))NR^(f)R^(g); NR^(e)C(NH)NR^(f)R^(g) or    —NR^(e)C(NR^(f))R^(g);-   each R^(e), R^(f) and R^(g) independently represents hydrogen;    —R^(h); —C₁₋₆ straight- or branched-chain alkyl unsubstituted or    substituted with one to four R^(i) groups;-   or R^(e) and R^(f) taken together with any intervening atoms    represent a 4-6 membered saturated ring optionally interrupted by    one to three of O, S, —C(O)— or NR^(g) with R^(g) as defined above,    said ring being unsubstituted or substituted with one to four R^(i)    groups;-   each R^(i) independently represents halo; —CN; —NO₂; phenyl;    —NHSO₂R^(h); —OR^(h), —SR^(h); —N(R^(h))₂; —N⁺(R^(h))3;    —C(O)N(R^(h))²; SO₂N(R^(h))₂; heteroaryl; heteroarylium; —CO₂R^(h);    —C(O)R^(h); —OCOR^(h); NHCOR^(h); guanidinyl; carbamimidoyl or    ureido;-   each R^(h) independently represents hydrogen, a —C₁₋₆ straight or    branched-chain alkyl group, a —C₃-C₆ cycloalkyl group or phenyl, or    when two R^(h) groups are present, said R^(h) groups may be taken in    combination and represent a 4-6 membered saturated ring, optionally    interrupted by one or two of O, S, SO₂, —C(O)—, NH and NCH₃;-   Q is selected from the group consisting of

wherein.

-   a and b are 1, 2 or 3;-   L⁻ is a pharmaceutically acceptable counterion;-   α represents O, S or NR^(s);-   β, δ, λ, μ, and σ represent CR^(t), N or N⁺R^(s), provided that no    more than one of β, δ, λ, μ, and σ is N⁺R^(s);-   each R^(s) independently represents hydrogen; phenyl or C₁₋₆    straight- or branched-chain alkyl, unsubstituted or substituted with    one to four R^(i) groups;-   each R^(t) independently represents hydrogen; halo; phenyl; —CN;    —NO₂; —NR^(u)R^(v); —OR^(u); —SR^(u); —CONR^(u)R^(v); —COOR^(h);    —SOR^(u); —SO₂R^(u); —SO₂NR^(u)R^(v); —NR^(u)SO₂R^(v); —COR^(u);    —NR^(u)COR^(v); —OCOR^(u); —OCONR^(u)R^(v); —NR^(u)CO₂R^(v);    —NR^(u)CONR^(v)R^(w); —OCO₂R^(v); —C₁₋₆ straight- or branched-chain    alkyl, unsubstituted or substituted with one to four R^(i) groups;-   R^(u) and R^(v) represent hydrogen or —C₁₋₆ straight- or    branched-chain alkyl, unsubstituted or substituted with one to four    R^(i) groups;-   or R^(u) and R^(v) together with any intervening atoms represent a    4-6 membered saturated ring optionally interrupted by one or more of    O, S, NRW or —C(O)—, said ring being unsubstituted or substituted    with one to four R^(i) groups;-   each R^(w) independently represents hydrogen; —C₁₋₆ straight- or    branched-chain alkyl, unsubstituted or substituted with one to four    R^(i) groups; C₃₋₆ cycloalkyl optionally substituted with one to    four R^(i) groups; phenyl optionally substituted with one to four    R^(i) groups, or heteroaryl optionally substituted with 1-4 R^(i)    groups; or R^(h) and R^(w) taken together with any intervening atoms    represent a 5-6 membered saturated ring, optionally interrupted by    one or two of O, S, SO₂, NH or NCH₃;-   R^(x) represents hydrogen or a C₁₋₈ straight- or branched chain    alkyl, optionally interrupted by one or two of O, S, SO, SO₂,    NR^(w), N⁺R^(h)R^(w), or —C(O)—, said chain being unsubstituted or    substituted with one to four of halo, CN, NO₂, OR^(w), SR^(w),    SOR^(w), SO₂R^(w), NR^(h)R^(w), N⁺(R^(h))₂R^(w), —C(O)—R^(w),    C(O)NR^(h)R^(w), SO₂NR^(h)R^(w), CO₂R^(w), OC(O)R^(w),    OC(O)NR^(h)R^(w), NR^(h)C(O)R^(w), NR^(h)C(O)NR^(h)R^(w), or a    phenyl or heteroaryl group which is in turn optionally substituted    with from one to four R^(i) groups or with one to two C₁₋₃ straight-    or branched-chain alkyl groups, said alkyl groups being    unsubstituted or substituted with one to four R^(i) groups;-   R^(y) and R^(z) represent hydrogen; phenyl; —C₁₋₆ straight or    branched chain alkyl, unsubstituted or substituted with one to four    R^(i) groups, and optionally interrupted by O, S, NR^(w),    N⁺R^(h)R^(w) or —C(O)—;-   or R^(x) and R^(y) together with any intervening atoms represent a    4-6 membered saturated ring optionally interrupted by O, S, SO₂,    NR^(w), N⁺R^(h)R^(w) or —C(O)—, unsubstituted or substituted with    1-4 R^(i) groups,-   and when R^(x) and R^(y) together represent a 4-6 membered ring as    defined above, R^(z) is as defined above or R^(z) represents an    additional saturated 4-6 membered ring fused to the ring represented    by R^(x) and R^(y) taken together, optionally interrupted by O, S,    NR^(w) or —C(O)—, said rings being unsubstituted or substituted with    one to four R^(i) groups.

In one sub-embodiment, of Formula I,

-   R is lower alkyl;-   R¹ is lower alkyl;-   M is H or a group such that CO₂M represents a carboxylic acid    protected by a protecting group;-   P is hydroxyl or hydroxyl protected by a hydroxyl protecting group;-   Y¹ is C₁₋₆ straight- or branched-chain alkyl that are substituted or    unsubstituted with one to four R^(d) groups and with or without    saturation (double or triple bonds); and-   Y² is independently selected from —CN; —NR^(a)R^(b);    —C(O)NR^(a)R^(b); —C(O)OR^(h); —NR^(a)SO₂R^(b); —C(O)R^(a) and    -A-(CH₂)_(n)-Q;-   wherein A represents —CH₂—;-   n represents an integer 0, 1, 2 or 3;-   each R^(a) and R^(b) independently represent hydrogen, —C₁₋₆    straight- or branched-chain alkyl, unsubstituted or substituted with    one to four R^(d) groups, or —C₃₋₇ cycloalkyl, unsubstituted or    substituted with one to four R^(d) groups; or R^(a) and R^(b) taken    together with any intervening atoms represent a 4-6 membered    saturated ring optionally interrupted by one or more of O, S,    NR^(c), with R^(c) as defined above, or —C(O)—, said ring being    unsubstituted or substituted with one to four R^(i) groups;-   each R^(d) independently represents halo; —CN; —NO₂; —NR^(e)R^(f);    —OR^(g); —SR^(g); —CONR^(e)R^(f); —COOR^(g); —SOR^(g); —SO₂R^(g);    —SO₂NR^(e)R^(f); —NR^(e)SO₂R^(f); —COR^(e); —NR^(e)COR^(f);    —OCOR^(e); —OCONR^(e)R^(f)l NR^(e)CONR^(f)R^(g); —NR^(e)CO₂R^(h);    —OCO₂R^(h); —C(NR^(e))NR^(f)R^(g); NR^(e)C(NH)NR^(f)R^(g) or    —NR^(e)C(NR^(f))R^(g);-   each R^(e), R^(f) and R^(g) independently represents hydrogen;    —R^(h); —C₁₋₆ straight- or branched-chain alkyl unsubstituted or    substituted with one to four R^(i) groups;-   each R^(i) independently represents halo; —CN; —NO₂; phenyl;    —NHSO₂R^(h); —OR^(h), —SR^(h); —N(R^(h))₂; —N⁺(R^(h))3;    —C(O)N(R^(h))₂; SO₂N(R^(h))₂; heteroaryl; heteroarylium; —CO₂R^(h);    —C(O)R^(h); —OCOR^(h); NHCOR^(h); guanidinyl; carbamimidoyl or    ureido;-   each R^(h) independently represents hydrogen, a —C₁₋₆ straight or    branched-chain alkyl group, a —C₃-C₆ cycloalkyl group or phenyl, or    when two R^(h) groups are present, said R^(h) groups may be taken in    combination and represent a 4-6 membered saturated ring, optionally    interrupted by one or two of O, S, SO₂, —C(O)—, NH and NCH₃;-   Q is selected from the group consisting of:

wherein.

-   a and b are 1, 2 or 3;-   L⁻ is a pharmaceutically acceptable counterion;-   α represents O, S or NR^(s);-   β, δ, λ, μ, and σ represent CR^(t), N or N⁺R^(s), provided that no    more than one of β, δ, λ, μ, and σ is N⁺R^(s);-   each R^(s) independently represents hydrogen; phenyl or C₁₋₆    straight- or branched-chain alkyl, unsubstituted or substituted with    one to four R^(i) groups;-   each R^(t) independently represents hydrogen; halo; phenyl; —CN;    —NO₂; —NR^(u)R^(v); —OR^(u); —SR^(u); —CONR^(u)R^(v); —COOR^(h);    —SOR^(u); —SO₂R^(u); —SO₂NR^(u)R^(v); —NR^(u)SO₂R^(v); —COR^(u);    —NR^(u)COR^(v); —OCOR^(u); —OCONR^(u)R^(v); —NR^(u)CO₂R^(v);    —NR^(u)CONR^(v)R^(w); —OCO₂R^(v); —C₁₋₆ straight- or branched-chain    alkyl, unsubstituted or substituted with one to four R^(i) groups;-   R^(u) and R^(v) represent hydrogen or —C₁₋₆ straight- or    branched-chain alkyl, unsubstituted or substituted with one to four    R^(i) groups;-   each R^(w) independently represents hydrogen; —C₁₋₆ straight- or    branched-chain alkyl, unsubstituted or substituted with one to four    R^(i) groups.

In another embodiment, Q is selected from

wherein a, b, L⁻, R^(x), R^(y) and R^(z) are as originally defined.

In one particular embodiment of the present invention, the carbapenem ofthe formula II,

or pharmaceutically acceptable salts and/or prodrugs thereof isprovided, whereinR¹ is hydrogen or methyl,P is hydrogen or hydroxyl, andY¹ is H or R², wherein R² is H or C₁-C₄ alkyl and Y² is —(CH₂)_(n)-A,wherein n=0-4 and A is —CN, —OR², —SR², —N(R²)₂, CO₂R², CO)—N(R²),—CO)—NR²SO₂N(R²)₂, NR²SO₂N(R²)₂, NH—C(═NR²)—N(R²)₂, —S—C(═NR²)—N(R²)₂,wherein each R² is H or C₁-C₄ alkyl.

In another particular embodiment of the present invention, thecarbapenem of the formula III,

or pharmaceutically acceptable salts and/or prodrugs thereof isprovided, whereinR¹ is hydrogen or methyl,P is hydrogen or hydroxyl, andY is —(CH₂)_(n)-A, wherein n=0-4 and A is —CN, —OR², —SR², —N(R²)₂,CO₂R², C(═O)—N(R²), —C(═O)—NR²SO₂N(R²)₂, NR²SO₂N(R²)₂,NH—C(═NR²)—N(R²)₂, —S—C(═NR²)—N(R²)₂, wherein each R² is H or C₁-C₄alkyl.

In another particular embodiment of the present invention, thecarbapenem of the formula IV,

or pharmaceutically acceptable salts and/or prodrugs thereof isprovided, whereinR¹ is H or methyl,P is hydrogen or hydroxyl,n=0-4, andB is —OR², —SR², —N(R²)₂, CO₂R², CO—N(R²), —CO—NR²SO₂N(R²)₂,NR²SO₂N(R²)₂, NH—C(═NR²)—N(R²)₂, —S—C(═NR²)—N(R²)₂, wherein each R² is Hor C₁-C₄ alkyl, independently.

In one subembodiment of Formula (IV), P′ is hydroxyl, R¹ is methyl, n=0,1, and B is CONH₂, —CONHSO₂NH₂, NHSO₂NH₂, NH—C(═NH)—NH₂, —S—C(═NH)—NH₂.

In another particular embodiment of the present invention, thecarbapenem of the formula V,

or pharmaceutically acceptable salts and/or prodrugs thereof isprovided, whereinR¹ is hydrogen or methyl,P is hydrogen or hydroxyl,Z is —O, —S, —N(R²), and,each Y³ and Y⁴ independently represents hydrogen or —(CH₂)_(n)-A,wherein n=0-4 and A=—N(R²)₂, —CO₂R², —CO—N(R²)₂, —CO—NR²SO₂N(R²)₂,NR²SO₂N(R²)₂, NH—C(═NR²)—N(R²)₂, —S—C(═NR²)—N(R²)₂, wherein each R² is Hor C₁-C₄ alkyl, independently.

In one subembodiment of Formula (V), P′ is hydroxyl, R¹ is methyl, Z isNH, and, each Y³ and Y⁴ independently represents hydrogen or—(CH₂)_(n)-A, wherein n=0-2 and A=—N(R²)₂, —CON(R²)₂, —CO—NHSO₂NH₂,NHSO₂NH₂, wherein R²=hydrogen or methyl.

In certain subembodiments, the compounds of the invention are thosecompounds shown in FIG. 2.

In certain subembodiments, the compound is compound 9:

or a pharmaceutically acceptable salt or prodrug thereof.

In certain subembodiments, the compound is compound 12:

or a pharmaceutically acceptable salt or prodrug thereof.

In certain subembodiments, the compound is compound 15:

or a pharmaceutically acceptable salt or prodrug thereof.

In certain subembodiments, the compound is compound 18:

or a pharmaceutically acceptable salt or prodrug thereof.

In certain subembodiments, the compound is compound 21:

or a pharmaceutically acceptable salt or prodrug thereof.

In certain subembodiments, the compound is compound 25:

or a pharmaceutically acceptable salt or prodrug thereof.

In certain subembodiments, the compound is compound 28:

or a pharmaceutically acceptable salt or prodrug thereof.

In certain subembodiments, the compound is compound 32:

or a pharmaceutically acceptable salt or prodrug thereof.

In certain subembodiments, the compound is compound 34:

or a pharmaceutically acceptable salt or prodrug thereof.

In certain subembodiments, the compound is compound 36:

or a pharmaceutically acceptable salt or prodrug thereof.

In certain subembodiments, the compound is compound 39:

or a pharmaceutically acceptable salt or prodrug thereof.

Process of Making Compounds

The compounds of the present invention can be prepared using the generalprocess outlined in Scheme 1, below, such as from carbapenemintermediate 5 illustrated in FIG. 3. Reagents and substrates usedherein can be either purchased, or synthesized according to knownprocedures.

The present invention also is directed to an efficient synthetic routeto β-methyl carbapenems from available precursors with the option ofintroducing functionality as needed. The process of synthesis isapplicable to a wide range of oxygen and nitrogen linkers, as well asother heteroatom linkers, such as sulfur and phosphorus. The carbapenemsmade according to the present invention can also be used as syntheticintermediates in the preparation of a variety of other β-methylcarbapenem analogs, as well as additional derivatives obtained bysubsequent functional group manipulations.

The invention also provides intermediates disclosed herein that areuseful in the preparation of compounds of the present invention as wellas synthetic methods for preparing the compounds of the invention.

In one embodiment of the invention, the carbapenem intermediate issynthesized using Scheme 1, which is shown in greater detail in FIG. 3.

This carbapenem intermediate, containing an activated —O(CO)₂R′, whereinR′ is an alkyl, such as —O(CO)₂(i-Bu), to achieve coupling of thecarbapenem to the heteroaromatic side chain, including a oxygen ornitrogen moiety, to produce a β-methyl carbapenem.

In one embodiment, a process for synthesizing a compound represented byformula I.

or a pharmaceutically acceptable salt thereof is provided.

The present invention also is directed to methods of efficient synthesisof β-methyl carbapenems from available precursors with the option ofintroducing functionality as needed. Therefore, in one embodiment, thecarbapenems are synthesized using the process of the present invention.

In one embodiment, the method for preparing β-methyl carbapenemsincludes:

-   a) preparing or obtaining a carbapenem intermediate of the structure    (A), for example using the process of Scheme 1

-   -   wherein    -   P, R and R¹ are as defined above;    -   P′ is a suitable carboxyl protecting group; and    -   R′ is an alkyl or substituted alkyl; and then

-   b) coupling the compound of structure (A) with a moiety with a free    hydroxyl, such as an aromatic alcohol or a heteroaromatic alcohol,    or a mono- or di-substituted amine, such as a aromatic amine or    heteroaromatic amine, to obtain an β-methyl carbapenem; and then

-   c) optionally deprotecting the β-methyl carbapenem, if necessary.

In one illustrative embodiment, the carbapenem intermediate (A), is thefollowing compound (A*).

The selection of reaction conditions should take into account the easeof substitution of the —O(CO₂)R in the carbapenem intermediate to formthe desired carbapenem.

The process of synthesis is applicable to a wide range of oxygen andnitrogen linkers, as well as other heteroatom linkers, such as sulfurand phosphorus. The carbapenems made according to the present inventioncan also be used as synthetic intermediates in the preparation of avariety of other β-methyl carbapenem analogs, as well as additionalderivatives obtained by subsequent functional group manipulations.

The invention also provides intermediates disclosed herein that areuseful in the preparation of compounds of the present invention as wellas synthetic methods for preparing the compounds of the invention.

Suitable solvents for carrying out the processes of the presentdisclosure are inert organic solvents, including but are not limited to,alcohols, aldehydes, amides, ethers, esters, halogenated solvents,hydrocarbons, glycols and glycol ethers, ketones, nitriles, and numerousother solvents common in chemical processes, as well as mixtures of suchsolvents. These inert solvents can be used alone or in combination, andcan be miscible or immiscible with each other, with the proviso onlythat the compounds of interest are at least partially soluble in thesolvent or solvents used. In the instance of using an immiscible, or2-phase, solvent system, the process can also include the addition of aphase-transfer agent. Suitable phase-transfer agents are known in theart, such as those described in Sasson, et al., Handbook of PhaseTransfer Catalysis, Kluwer Academic Publishers, 1997.

In one embodiment, the solvent is DMF, the reaction is carried out atabout room temperature, using 5 mol eq. of Pd₂ dba₃*CHCl₃(tris(dibenzylideneaceton)-dipalladium(0)-chloroform adduct), 30 mol eq.of either P(OEt)₃, no acid or base. In certain cases, 0.5 eq.2,6-lutidine can be added to increase the rate of reaction. In certainembodiments, 0.5 eq. PTSA are added.

In one embodiment, the reaction is run with dppb(1,4-bis(diphenylphosphine)butane) or triethyl phosphate.

In one embodiment, the reaction is run in the absence of a base. Inanother embodiment, the reaction is carried out (run) in the absence ofan acid. In a separate embodiment the reaction is carried out in2,6-lutidine or p-toluenesulfonic acid monohydrate.

For the purpose of the present invention, the inert organic solventssuitable for use in preparing the compounds described and claimed hereininclude but are not limited to aromatic solvents, such as benzene,toluene, chloro benzene, styrene, tetraline, biphenyl, and xylenes;ether solvents, such as diethyl ether, n-butyl ether, methyl tert-butylether, tetrahydrofuran (THF), and 1,4-dioxane; halogenated solvents,such as chloroform, bromoform, carbon tetrachloride, dichloromethane,dichloroethane, trichloroethane, dichlorobenzene, and chlorobenzene;alcohols, including C₁-C₁₀ alkanols, which can be linear, branched, orcyclic, and may be saturated or unsaturated, including methanol,ethanol, 2-propanol, butanol and hexanol; C₁-C₁₀ hydrocarbon solvents,which can be linear, branched, or cyclic, and may be saturated orunsaturated, including hexane, heptane, cyclohexane, cyclohexene, andpentane; ester and ketone solvents, such as acetone, ethyl acetate,isopropyl acetate, methylbutyl ketone (2-hexanone), methyl ethyl ketone(MEK), methylisobutyl ketone (MIBK), methyl n-butyl ketone (MBK), methylisopropyl ketone, and cyclohexanone; and nitrogen-containing solvents,including acetonitrile, nitromethane, N,N-dimethylformamide (DMF),dimethylacetamide (DMA), hexamethylphosphoramide (HMPA),N-methylpyrrolidinone (NMP), N,N′-dimethylpropylene urea (DMPU),1,3-dimethyl-2-oxohexahydropyrimidine, and N-ethylpyrrolidinone.

Bases suitable for use in conducting certain of the synthetictransformations described and claimed herein include but are not limitedto carbonates, including alkali metal carbonates and bicarbonates, suchas sodium carbonate, sodium bicarbonate, potassium carbonate, rubidiumcarbonate, and cesium carbonate; alkaline earth metal carbonates, suchas magnesium carbonate, calcium carbonate, and strontium carbonate;hydroxides, such as sodium hydroxide and potassium hydroxide; andtransition metal bases, such as zinc hydroxide. Also suitable for use asbases in the transformations described herein are organic bases,including but not limited to triethylamine (TEA); diethylamine;diisopropylamine; N,N-diisopropylethylamine (DIPEA or DIEA, also knownas Hunig's base); dimethylamine; benzylamine; 4-dimethylaminopyridine(DMAP); ureas, such as tetramethylurea (TMU); pyridine; 2,6-lutidine;imidazole; pyrrole; diphenylamine; tri-n-propylamine; cyclohexylamine;triphenylamine; pyrrolidine; ureas, such as tetramethylurea (TMU); andpiperidine.

As defined above, when a functional group is termed to be “protected”with a “protecting group” (herein represented by the letter designation,P), this means that the group is chemically modified to precludeundesired side reactions at the protected site. Suitable compounds foruse with the compounds of the present invention will be recognized fromthe present application, and include those included in such standardreference texts known to those of skill in the art as Greene, T. W. andWuts, P. G. M., “Protective Groups in Organic Synthesis, Third Edition”,Wiley Interscience, New York (1999). Examples of suitable protectinggroups include but are not limited to silyl protecting groups, includingtri-C₁₋₆ alkyl silyl groups (e.g., trimethylsilyl and triethylsilyl),diphenyl siloxy groups (e.g., t-butyldiphenylsilyl (TBDPS)), C₁₋₆ alkylsilyloxy groups (e.g., tert-butyldimethylsilyl (TBDMS)), substituted andunsubstituted benzyl groups (e.g., benzyl, benzyloxycarbonyl,o-nitrobenzyloxycarbonyl, p-nitrobenzyloxycarbonyl), carbonyls, such as2,2,2-trichloroethyloxycarbonyl (Troc), allyloxycarbonyl (Alloc), andfluorenylmethyloxycarbonyl (Fmoc).

The processes of preparing the compounds of the present invention aresuitably conducted at a temperature in a range of from about −78° C. tothe boiling point of the reaction medium or solvent (e.g., from about−78° C. to about 200° C.), and are typically conducted at a temperaturein a range of from about −50° C. to the boiling point of the reactionmedium or solvent. In one embodiment, the temperature is in a range offrom about −20° C. to the boiling point of the reaction medium orsolvent. In another embodiment, the temperature is in the range of fromabout −10° C. to the boiling point of the reaction medium or solvent.

The reactants used in the presently disclosed process can be added tothe reaction vessel (also referred to herein as the reaction “pot”, or“round bottom”) concurrently, either together or separately, or they canbe added sequentially in either order.

Method of Treatment

The present invention also provides a method of preventing or treating abacterial infection, in a host, for example an animal, and typically ahuman, including administering a therapeutic amount of a compound of thepresent invention, or a pharmaceutically acceptable salt and/or prodrugtherein, optionally in a pharmaceutically acceptable carrier or diluentwhere the bacterial infection is due to a gram-negative bacteria. In oneembodiment, the bacterial infection is a drug resistant and/ormultiple-drug resistant bacterial infection.

The invention also provides a compound of the present invention for usein medical therapy.

The present invention also provides a use of a therapeutic amount of acompound of the present invention, or a pharmaceutically acceptable saltand/or prodrug therein, optionally in a pharmaceutically acceptablecarrier or diluent, for preventing or treating a gram-negative bacterialinfection, in a host, such as an animal, and typically a human.

The distinctive feature of gram-negative bacteria is the presence of adouble membrane surrounding each bacterial cell. Although all bacteriahave an inner cell membrane, gram-negative bacteria have a unique outermembrane. This outer membrane excludes certain drugs and antibioticsfrom penetrating the cell, partially accounting for why gram-negativebacteria are generally more resistant to antibiotics than aregram-positive bacteria. The pathogenic capability of gram-negativebacteria is usually associated with certain components of their cellwalls, particularly the lipopolysaccharide (endotoxin) layer. The outermembrane of gram-negative bacteria is rich in lipopolysaccharide. Ifgram-negative bacteria enter the bloodstream, lipopolysaccharide cantrigger a cascade of events, including high fever and a drop in bloodpressure. Unlike Gram-positive bacteria, which assume a violet color inGram staining, Gram negative bacteria incorporate the counterstainrather than the primary stain. Because the cell wall of Gram(−) bacteriais high in lipid content and low in peptidoglycan content, the primarycrystal-violet escapes from the cell when the decolorizer is added. Mostenteric (bowel related) illnesses can also be attributed to this groupof bacteria.

Examples of gram-negative bacteria include Aeromonas sp., Acinetobactersp. such as Acinetobacter baumannii (or A. calcoaceticus),Actinobacillus actinomycetemcomitans, Bacteroides sp. such asBacteroides fragilis, Bartonella, Bdellovibrio spp., Bordetellapertussis, Brucella sp., Burkholderia cepacia, Burkholderia,pseudomallei, Campylobacter sp., Capnocytophaga sp., Cardiobacteriumhominis, Chlamydia trachomatis, Citrobacter sp., Eikenella corrodens,Enterobacter sp., Escherichia coli, Francisella tularensis,Flavobacterium sp., Fusobacterium sp., Helicobacter pylori, Haemophilusinfluenzae, Haemophilus ducreyi, Klebsiella spp. such as Klebsiellapneumoniae, Kingella kingae, Legionella spp. such as Legionellapneumophila, Moraxella catarrhalis, Morganella, Neisseria gonorrhoeae,Neisseria meningitidis, Pasteurella pestis, Pasteurella multocida,Plesiomonas shigelloides, Prevotella sp., Proteus spp., Providencia,Pseudomonas spp. such as Pseudomonas aeruginosa, Salmonella spp. such asSalmonella enteriditis and Salmonella typhi, Serratia marcescens,Shigella spp., Vibrio cholerae, Vibrio parahaemolyticus, Vibriovulnificus, Veillonella sp., Xanthomonas maltophilia or Stenotrophomonasmaltophila, Yersinia pestis, Yersinia enterocolitica. Additionally, Someorganisms simply tend not to be well differentiated by gram staining,despite any known phylogenetic affiliation with the gram-negatives orgram-positives. Rickettsia prowazekii, Rickettsia rickettsii andTreponema pallidum. Chlamydias are small, gram-negative,peptidoglycan-less cocci that are obligate intracellular parasites ofanimals. Spirochetes are chemoheterotrophic bacteria whose cells aretightly coiled or resemble a stretched spring with gram-negative-likecell envelopes. Spirochetes include Spirillum minus, Borreliaburgdorferi (Lyme disease), Leptospira spp. (leptospirosis) andTreponema pallidum (syphilis). Rickettsias and actinomycetes are alsogram-negative pleomorphic bacilli and coccobacilli that are obligateintracellular parasites of eucaryotes transmitted generally by insectsand ticks.

The present invention also provides a use of a therapeutic amount of acompound of the present invention, or a pharmaceutically acceptable saltand/or prodrug therein, optionally in a pharmaceutically acceptablecarrier or diluent, in the manufacture of a medicament for preventing ortreating a gram-negative bacterial infection, in a host, such as ananimal, and typically a human.

The invention also includes methods of inhibiting bacterial infection ina host. Inhibition of bacterial replication or treatment of an infectionin a cell can be measured by showing a reduction in bacterialreplication in a cell to a level lower than the level in an otherwiseidentical cell, which was not administered the compound of theinvention. The reduction can be by about 80%, 85%, 90%, 95%, about 99.9%or more. The level of bacterial replication in a cell can be assessed byany known methods. For example, the level of bacterial replication in acell can be assessed by evaluating the number of bacterial particles oramount of a bacterial component, such as a bacterial protein, abacterial enzyme, or bacterial nucleic acid, in the cell or in fluid ordebris associated with the cell. The number of infectious bacteria in acell can be evaluated, for example, in a plaque assay. The level of abacterial component such as a bacterial protein or enzyme in a cell canbe evaluated using standard analytical techniques of proteinbiochemistry, such as, for example, using an activity assay for abacterial enzyme, or using Western blotting or quantitative gelelectrophoresis for a bacterial protein. Bacterial nucleic acid levelsin a cell can be evaluated using standard analytical techniques such asNorthern blotting and Southern Blotting or quantitation by polymerasechain reaction (PCR).

Combination and Alternation Therapies

In one embodiment of the invention, one or more therapeutic agents,including particularly antimicrobial agents such as antibiotic agentsthat are effective against gram negative bacteria, can be used incombination and/or alternation with the compound/composition of thepresent invention to achieve a additive and/or synergistic therapeuticeffect.

The active compounds can be administered in combination, alternation orsequential steps with another anti-bacterial agent. In combinationtherapy, effective dosages of two or more agents are administeredtogether, whereas in alternation or sequential-step therapy, aneffective dosage of each agent is administered serially or sequentially.The dosages given will depend on absorption, inactivation and excretionrates of the drug as well as other factors known to those of skill inthe art. It is to be noted that dosage values will also vary with theseverity of the condition to be alleviated. It is to be furtherunderstood that for any particular subject, specific dosage regimens andschedules should be adjusted over time according to the individual needand the professional judgment of the person administering or supervisingthe administration of the compositions. In some embodiments, ananti-bacterial agent that exhibits an EC₅₀ of 10-15 μM or less, ortypically less than 1-5 μM.

In one particular embodiment, the combination includes a β-lactamaseinhibitor, such as clavulanic acid, which has been used as in thedelivery of prophylactic amounts of antibiotics in patients. AlthoughClavulanic acid does have some degree of bacterial activity, itsprincipal role is as a beta-lactamase inhibitor. Clavulanic acid has asimilar structure to the beta-lactam antibiotics but binds irreversiblyto the beta-lactamase enzymes. Used in combination with the beta-lactamantibiotics, it has become one of the most prescribed antibiotics in thewestern world prolonging the effective life of antibiotics such asAmpicillin (as in GSK's Augmentin®).

It is possible that drug-resistant variants of bacteria can emerge afterprolonged treatment with an anti-bacterial agent. The efficacy of a drugagainst the bacterial infection can be prolonged, augmented, or restoredby administering the compound in combination or alternation with asecond, and perhaps third, anti-bacterial agent, for example with adifferent site of activity than the principle drug. Alternatively, thepharmacokinetics, biodistribution or other parameter of the drug can bealtered by such combination or alternation therapy.

Suitable antibiotic agents are disclosed, e.g. in Physician's Desk 30Reference (PDR), Medical Economics Company (Montvale, N.J.), (53rd Ed.),1999; Mayo Medical Center Formulary, Unabridged Version, Mayo Clinic(Rochester, Minn.), January 1998; Merck Index An Encyclopedia ofChemicals, Drugs and Biologicals, (11th Ed.), Merck & Co., Inc. (Rahway,N.J.), 1989; University of Wisconsin Antimicrobial Use Guide,http://www.medsch.wisc.edu/clinsci/5amcg/amcg.html; Introduction on theUse of the Antibiotics Guideline, of Specific Antibiotic Classes, ThomasJefferson University,http://jeffiine.tju.edu/CWIS/OAC/antibiotics_guide/intro.html; andreferences cited therein.

Nonlimiting examples of agents that can be used in combination oralternation with the compounds of the invention include:aminoglycosides, β-lactam antibiotics, cephalosporius, macrolides,miscellaneous antibiotics, penicillins, tetracyclines, antifungals,antimalarial agents, antituberculosis agents, antibacterials,leprostatics, miscellaneous anti-infectives, quinolones, sulfonamides,urinary anti-infectives, nasal antibiotics, ophthalmic antibiotics,ophthalmic antibacterials, ophthalmic quinalones, ophthalmicsulfonamides, skin and mucous membrane antibiotics, skin and mucousmembrane antifungals, skin and mucous membrane antibacterials, skin andmucous membrane miscellaneous anti-infectives, skin and mucous membranescabicides and pedulicides, skin and mucous membrane antineoplasts,nitrofurans and oxazolidinones.

Specific compounds include, for example, Amikacin (amikacin sulfate);Craramyein (gentamicin sulfate); Nebcin (tobramycin sulfate); Netromycin(netilmicin sulfate); Streptomycin Sulfate; and TOBI (tobramycin),Azactam (aztreonam); Cefotan (cefotetan); Lorabid (loracarbef); Mefoxin(cefoxitin); Merrem (meropenem); and Primaxin (imipenem and cilastatinfor injectable suspension); Ancef (cefazolin); Ceclor (cefaclor); Cedax(ceffibuten); Cefizox (ceffizoxime sodium); Cefobid (cefoperazonesodium); Ceftin (cefuroxime axetil); Cefzil (cefprozil); Ceptaz(ceftazidime); Claforan (cefotaxime); Duricef (cefadroxil monohydrate);Fortaz (ceftazidime); Keflex (cephalexin); Keftab (cephalexin HCl);Kefurox (cefuroxime); Kefzol (cefazolin); Mandol (cefamandole nafate);Maxipime (cefepime HCl); Monocid (cefonicid sodium); Omnicef (cefdinir);Rocephin (ceftriaxone); Suprax (cefixime); Tazicef (ceftazidime);Tazidime (ceftazidime); Vantin (cefpodoxime proxetil); and Zinacef5(cefuroxime); Biaxin (clarithromycin); Dynabac (dirithromycin); E.E.S.200 (Erythromycin Ethylsuccinate); E.E.S. 400 (ErythromycinEthylsuccinate); Ery-Ped 200 (Erythromycin Ethylsuccinate); EryPed 400(Erythromycin Ethylsuccinate); Ery-Tab (Erythromycin delayed-releasetablets); Erythrocin Stearate (Erythromycin stearate); Ilosone(erythromycinestolate); PCE Dispertab (erythromycin particles intablets); Pediazole (erythromycin ethylsuccinate and sulfisoxazoleacetyl for oral suspension); Tao (troleandomycin); Zithromax(azithromycin); and Erythromycin; Cleocin HCl (clindamycinhydrochloride); Cleotin Phosphate (elindamycin phosphate); Coly-Mycin M(colistimethate sodium); and Vancocin HCl (vancomycin hydrochloride);Amoxil (amoxicillin); Augmentin (amoxicillin/clavulanate potassium);Bicillin C-R 900/300 (Penicillin G benzathine and Penicillin G procainesuspension); Bicillin C-R (Penicillin G benzathine and Penicillin Gprocaine suspension); Bicillin L-A (Penicillin G benzathine suspension);Geoeillin (carbencillin indanyl sodium); Mezlin (sterilemezlocillinsodium); Omnipen (ampicillin); Pen-Vee K (penicillin Vpotassium); Pfizerpen (penicillin G potassium); Pipracil (piperacillinsodium); Speetrobid (bacampicillin-HCl); Ticar (tiearcillin disodium);Timentin (ticarcillin disodium and clavulanate potassium); Unasyn(ampicillin sodium/sulbactam sodium); Zosyn (piperacillin sodium andtazobactam sodium); and Dicloxacillin Sodium; Achromycin V (tetracyclineHCl); Declomycin (demeclo-cycline HCl); Dynacin (minocylcine HCl);Minocin (minocycline hydrochloride); Monodox (Doxycycline monohydratecapsules); Terramycin (oxytetracyline); Vectrin (minocyclinehydrochloride); Vibramycin Calcium (doxycycline sodium); VibramycinHyclate (doxycycline hyclate); Vibramycin Monohydrate (doxycyclinemonohydrate); Vibra-Tabs (doxycycline-hydrate); Declomycin(demeclocycline HCl); Vibramycin (doxycycline); Dynacin (MinocylineHCl); Terramycin (oxytetracycline HCl); Achromycin V capsules5(tetracycline HCl); Linco-mycins; and Cleotin HCl (clindamycin HCl);Abelcet (amphotericin B lipid complex); AmBisome (amphotericin B);Amphotec (amphotericin B cholesterol sulfatecomplex); Ancobon(flucytosine); Diflucan (fluconazole); Fulvicin P/Gamma (ultramicrosizegriseofulvin); Fulvicin P/G 165 and 330 (ultramicrosize griseofulvin);Grifulvin V (griseofulvin); Gals-PEG (gxiseofulvin ultramicrosize);Lamisil (terbinafine hydrochloride); Nizoral (ketoconazole);Amphotericin B; Lotrimin (clotrimazole); Dapsone tablets (dapsone);Diflucan (fluconazole); Monistat-Derm cream (miconazole); Mycostalin Crc.am (nystatin); and Sporanox (itraconazole); Aralen hydrochloride(chloroquine HCl); Aralen phosphate (chloroquine phosphate); Dataprim(pyrimethamine); Ladam (mefloquine HCl); and Plaquenil(hydroxychloroqnine sulfate); Capastat sulfate (capreomycinsulfate);Myambutol (ethambutol hydrochloride); Mycobutin (rifabutin capsules);Nydrazid (isoniazid injection); Paser (aminosalicylic acid); Prifiin(rifapentine); Pyrazinamide tablets (pyrazinamide); Rifadin (rifampincapsules); Rifadin IV (rifampin for injection); Rifamate (rifampin andisoniazid); Rifater (rifampin, isoniazid and pyrazinamide); Seromycin(cycloserine capsules); Streptomycin-Sulfate; Tice BCG (BCG vaccine);Cycloserine (seromycin capsules); Urised (Methenamine); and Trecator-SC(ethionamide tablets); Alferon N (interferon alfa-n3); Crixivan(indinavir sulfate); Cytovene (ganciclovir); Cytovene-IV (ganciclovirsodium); Epivir (lamivudine); Famvir (famciclovir); Flumadine(rimantadine HCl); Foscavir (foscarnet sodium); Hivid (zalcitabine);Intron A (interferon alfa-2b); Invirase (saquinavir mesylate); Norvir(ritonavir); Rebetron combination therapy, which contains Rebetrol(ribavirin) and Intron A (inteferon alfa-2b); Rescriptor (delavirdinemesylate); Retrovir (ziduvudine); Retrovir IV (ziduvudine); Symmetrel(amantadine HCl); Synagis (palivizumab); Valtrex (valacyclovir HCl);Videx (didanosine); Viracept (nelfinavir mesylate); Viramune(nevirapine); Virazole (ribavirin); Vistide (cidofovir); Zerit(stavudine (d4T)); Symmetrel Syrup (amantadine HCl); Combivir Tablets(lamiduvine); and Zovirax (acyclovir); Dapsone Tablets (dapsone);Daraprim (pyrimethamine); Flagyl 375 (metronidazole); Flagyl ER Tablets(metronidazole); Flagyl I.V. (metronidazole); Furoxone (furazolidone);Mepron (atovaquone); and Neutrexin (trimetrexate glucuronate); Cipro(ciprofloxacin HCl); Floxin (ofloxacin); Levaquin (levofloxacin);Mazaquin (lomefioxacin HCl); Noroxin (norfloxacin); Penetrex (enoxacin);Raxar (grepafloxacin HCl); Trovan (trovafloxacin mesylate); and Zagam(sparfloxacin); Bactrim.(trimethoprim and sulfamethoxazole); Bactrim DS(Irimethoprim and sulfamethoxazole double strength); Pediazole(erythromycin ethylsuccinate and sulfisoxazole acetyl); Septra(trimethoprim and sulfamethoxazole); Septra DS (trimethoprim andsulfamethoxazole); Co-Trimoxazole, Sulfadiazine, Battrim I.V. Infusion(sulfamethoxazole); Sulfapyridine and Pediazole (erythromycinethylsuccinate and sulfisoxazole acetyl); Furadantin (nitrofurantoin);Macrobid (nitrofurantoin monohydrate macrocrystals); Macrodantin(nitrofurantoin macrocrystals); Monurol Sachet (fosfomycintromethamine); NegGram Caplets (nalidixic acid); Septra (trimethoprimand sulfamethoxazole); Septra DS (trimethoprim and sulfamethoxazole);Urised (a combination of the antisepticsmethenamine, methylene blue,phenyl salicylate, benzoic acid and parasympatholytics (atropinesulfate) hyoscyamine); (oxytetracycline HCl, sulfamethizole andphenazopyridine HCl); (methenamine mandelate); Bactroban (mupirocin);Chloromycetin ophthalmic (chloramphenical); Cortisporin (neomycin andpolymyxin [3 sulfates and hydrocortisone acetate cream); Ilotycin(erythromycin ophthalmic ointment); NeoDecadron (neomycinsulfate-dexamethasone sodium phosphate); Polytrim (trimethoprim andpolythyxin [3 sulfate ophthalmic solution); Terra-Cortril(oxytetracycline HCl and hydrocortisone acetate); Terramycin(oxytetracycline); and TobraDex (tobramycin and dexamethasone ophthalmicsuspension and ointment); Vita-A ophthalmic ointment, (vidatabine);(norfloxacin ophthalmic solution; Ciloxan ophthalmic solution andointment (Ciprofloxacin HCl); and Ocuflox ophthalmic solution(ofloxacin), Blephamide ophthalmic ointment (sulfacetamide sodium andprednisolone acetate); and Blephamide ophthalmic suspension(sulfacetamide sodium and predrdsolone acetate); A/T/S (erythromycin);Bactroban (mupirocin); Benzamycin (erythromycin-benzoyl peroxide topicalgel); Betadine (povidone-odine); Cleotin T (clindamy cinphosphatetopical solution); Clindets (clindamycin phosphate pledgets);Cortispofin (neomycin, polymyxin B sulfates and hydrocortisone acetatecream); Emgel (erythromycin); Erycette (erythromycin topical solution);Garamycin (gentamicin sulfate); Klaron (sodium sulfacetamide lotion);Mycostatin (nystatin cream); Theramycin Z (erythromycin topicalsolution); T-Stat (erythromycin); Chloromycetin (chloramphenicolophthalmic ointment); Cortisporin (neomycin and polymyxin B sulfates,bacitracin zinc and hydrocortisone ophthalmic ointment); Ilotycin(erythromycin); NeoDeeadron (neomycin sulfate-dexamethasone sodiumphosphate); Polytrim (trimethoprim and polymyxin B sulfate);Terra-Cortril (oxytetracycline HCl and hydrocortisone acetate);Terramycin (oxytetracycline); Exelderm (sulconazole nitrate); Fungizone(amphotericin B oral suspension); Lamisil (terbinafine hydrochloridecream); Loprox (ciclopiroxolamine); Lotrimin (clotrimazole); Lotrisone(clotrimazole and betamethasone diproprionate); Mentax (butenafine HCl);Monistat-Denn (miconazole nitrate); Mycelex (clotrimazole); Mycostatin(nystatin); Naffin (nattifine HCl); Nizoral Ocetoconazole); Nystop(nystatin); Oxistat (oxiconazole nitrate); Selsun Rx (2.5% seleniumsulfide lotion); and Spectazole (econazole nitrate); Denavir(penciclovir cream); and Zovirax (acyclovir); Benzashave Coenzoylperoxide); Betadine (povidone-iodine); Betasept (chlorhexidinegluconate); Cetaphil (soap substitute); Clorpactin WCS-90 (sodiumoxychlorosene); Dapsone Tablets (dapsone); Desquam-E Coenzoyl peroxide);Desquam-X (benzoyl peroxide); Hibiclens (chlorhexidine gluconate);Hibistat (chlorhexidine gluconate); Impregon (tetrachlorosalicylanilide2%); MetroCream (metronidazole); MetroGel (metronidazole); Noritate(metronidazole); pHisoHex (hexachlorophene detergent cleanser);Sulfacet-R (sodium sulfacetamide 10% and sulfur 5%); Sulfamylon(materfide acetate); Tfiaz Coenzoyl peroxide); and Vanoxide-HC Coenzoylperoxide hydrocortisone); Acticin (permethrin); Elimite (permethrin);Eurax (crotamiton); Efudex (fluoro-uracil); Fluoroplex.

Pharmaceutical Compositions

Hosts, including humans can be treated by administering to the patientan effective amount of the active compound or a pharmaceuticallyacceptable prodrug or salt thereof in the presence of a pharmaceuticallyacceptable carrier or diluent. The active materials can be administeredby any appropriate route, for example, orally, parenterally,intravenously, intradermally, subcutaneously, or topically, in liquid orsolid form.

An optional dose of the compound for treatment of a bacterial (such as agram negative bacteria) infection is about 1 to 50 mg/kg, or 1 to 20mg/kg, of body weight per day, more generally 0.1 to about 100 mg perkilogram body weight of the recipient per day. The effective dosagerange of the pharmaceutically acceptable salts and prodrugs can becalculated based on the weight of the parent nucleoside to be delivered.If the salt or prodrug exhibits activity in itself, the effective dosagecan be estimated as above using the weight of the salt or prodrug, or byother means known to those skilled in the art.

Optionally, the active ingredient should be administered to achieve peakplasma concentrations of the active compound of from about 0.2 to 70 M,e.g., about 1.0 to 10 uM. This may be achieved, for example, by theintravenous injection of a 0.1 to 5% solution of the active ingredient,optionally in saline, or administered as a bolus of the activeingredient. The concentration of active compound in the drug compositionwill depend on absorption, inactivation and excretion rates of the drugas well as other factors known to those of skill in the art. It is to befurther understood that for any particular subject, specific dosageregimens should be adjusted according to the individual need and theprofessional judgment of the person administering or supervising theadministration of the compositions, and that the concentration rangesset forth herein are exemplary only and are not intended to limit thescope or practice of the claimed composition. The active ingredient maybe administered at once, or may be divided into a number of smallerdoses to be administered at varying intervals of time.

The compound is conveniently administered in unit any suitable dosageform, including but not limited to one containing 7 to 3000 mg, or 70 to1400 mg of active ingredient per unit dosage form. A dosage of 50-1000mg is optional.

The active compound can be administered in a pharmaceutically acceptablecarrier available in the art, and can be administered by a chosen routeof administration. Pharmaceutical compositions can be prepared,packaged, or sold in a variety of formulations which can be suitable forone or more routes of administration such as, for example, oral,intravenous, intramuscular, topical, subcutaneous, rectal, vaginal,parenteral, pulmonary, intranasal, buccal, ophthalmic, or another routeof administration. The active materials can be administered in liquid orsolid form. Other contemplated formulations include projectednanoparticles, liposomal preparations, resealed erythrocytes containingthe active ingredient, and immunologically-based formulations.

The active compound may be administered intravenously orintraperitoneally by infusion or injection. Solutions of the activecompound or its salts may be prepared in water or saline, optionallymixed with a non-toxic surfactant. Dispersions may be prepared inglycerol, liquid polyethylene glycols, triacetin, mixtures thereof, andin oils. Under ordinary conditions of storage and use, thesepreparations contain a preservative to prevent growth of microorganisms.

Pharmaceutical dosage forms suitable for injection or infusion mayinclude sterile aqueous solutions or dispersions or sterile powderscomprising the active ingredient which are adapted for theextemporaneous preparation of sterile injectable or infusible solutionsor dispersions, optionally encapsulated in liposomes. The ultimatedosage form is optionally sterile, fluid, and stable under conditions ofmanufacture and storage. The liquid carrier or vehicle may be a solventor liquid dispersion medium comprising, for example, water, ethanol, apolyol (for example, glycerol, propylene glycol, liquid polyethyleneglycols, and the like), vegetable oils, nontoxic glyceryl esters, andsuitable mixtures thereof.

For oral therapeutic administration, the active compound can be combinedwith one or more excipients and used in the form of ingestible tablets,buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers,and the like. Such compositions and preparations can contain at least0.1% (w/w) of active compound. The percentage of the compositions andpreparations can, of course, be varied, for example from about 0.1% tonearly 100% of the weight of a given unit dosage form. The amount ofactive compound in such therapeutically useful compositions is such thatan effective dosage level will be obtained upon administration.

The tablets, troches, pills, capsules, and the like may also contain oneor more of the following: binders, such as microcrystalline cellulose,gum tragacanth, acacia, corn starch, or gelatin; excipients, such asdicalcium phosphate, starch or lactose; a disintegrating agent, such ascorn starch, potato starch, alginic acid, primogel, and the like; alubricant, such as magnesium stearate or Sterotes; a glidant, such ascolloidal silicon dioxide; a sweetening agent, such as sucrose,fructose, lactose, saccharin, or aspartame; a flavoring agent such aspeppermint, methylsalicylate, oil of wintergreen, or cherry flavoring;and a peptide antibacterial agent, such as envuvirtide (Fuzeon™). Whenthe unit dosage form is a capsule, it can contain, in addition tomaterials of the above type, a liquid carrier, such as a vegetable oilor a polyethylene glycol. Various other materials may be present ascoatings or to otherwise modify the physical form of the solid unitdosage form.

In one embodiment, the active compounds are prepared with carriers thatwill protect the compound against rapid elimination from the body, suchas a controlled release formulation, including implants andmicroencapsulated delivery systems. Biodegradable, biocompatiblepolymers may be used, such as ethylene vinyl acetate, polyanhydrides,polyglycolic acid, collagen, polyorthoesters, and polylacetic acid.Methods for preparation of such formulations will be apparent to thoseskilled in the art. The materials may also be obtained commercially fromAlza Corporation.

Other formulations can also be developed. For example, the compounds canbe administered in liposomal suspensions (including liposomes targetedto infected cells with monoclonal antibodies to bacterial antigens).These may be prepared according to methods known to those skilled in theart, for example, as described in U.S. Pat. No. 4,522,811. For example,liposome formulations may be prepared in a variety of lipid(s) (such asstearoyl phosphatidyl ethanolamine, stearoyl phosphatidyl choline,arachadoyl phosphatidyl choline, and cholesterol).

A pharmaceutical composition of the invention may be prepared, packaged,or sold in a formulation suitable for rectal administration. Such acomposition may be in the form of, for example, a suppository, aretention enema preparation, and a solution for rectal or colonicirrigation. A pharmaceutical composition of the invention may also beprepared, packaged, or sold in a formulation suitable for vaginaladministration. Such a composition may be in the form of, for example, asuppository, an impregnated or coated vaginally-insertable material suchas a tampon, a douche preparation, or a solution for vaginal irrigation.

A pharmaceutical composition of the invention may be prepared, packaged,or sold in a formulation suitable for pulmonary administration via thebuccal cavity. Such a formulation may comprise dry particles whichcomprise the active ingredient and which have a diameter in the rangefrom about 0.5 to about 7 nanometers, or from about 1 to about 6nanometers. Such compositions are conveniently in the form of drypowders for administration, which can include particles wherein at least98% of the particles by weight have a diameter greater than 0.5nanometers and at least 95% of the particles by number have a diameterless than 7 nanometers. Typically least 95% of the particles by weighthave a diameter greater than 1 nanometer and at least 90% of theparticles by number have a diameter less than 6 nanometers. The activeingredient can also be in the form of droplets of a solution orsuspension, for example those that have an average diameter in the rangefrom about 0.1 to about 200 nanometers.

The formulations described herein as being useful for pulmonary deliveryare also useful for intranasal delivery of a pharmaceutical compositionof the invention. Another formulation suitable for intranasaladministration is a coarse powder comprising the active ingredient andhaving an average particle from about 0.2 to 500 micrometers.

A pharmaceutical composition of the invention may be prepared, packaged,or sold in a formulation suitable for ophthalmic administration. Fortopical administration, the present compounds can be applied in pureform, i.e., as a liquid. However, typically, the compounds areadministered to the skin as compositions or formulations, in combinationwith a dermatologically acceptable carrier. Useful solid carriersinclude finely divided solids such as talc, clay, microcrystallinecellulose, silica, alumina, and the like. Useful liquid carriers includewater, alcohols, glycols, and blends of two or more of these, in whichthe present compounds can be dissolved or dispersed at effective levels,optionally with the aid of non-toxic surfactants. Adjuvants such asfragrances and additional antimicrobial agents can be added to optimizeproperties for a given use. The resulting liquid compositions can beapplied using absorbent pads, used to impregnate bandages or otherdressings, or sprayed onto the affected area using pump-type or aerosolsprayers.

The compounds/compositions of the present invention are optionallyadministered in a controlled release formulation, which can be adegradable or nondegradable polymer, hydrogel or ganogel or otherphysical construct that modifies the bioabsorption, half life orbiodegradation of the active agent(s). The controlled releaseformulation can be a material that is painted or otherwise applied ontothe afflicted site, either internally or externally. In one embodiment,the invention provides a biodegradable bolus or implant. The controlledrelease formulation with appropriated selected imaging agent can be usedto coat a transplanted organ or tissue to prevent rejection. It canalternatively be implanted or otherwise applied near the site ofpotential infection.

Thickeners such as synthetic polymers, fatty acids, fatty acid salts andesters, fatty alcohols, modified celluloses, or modified mineralmaterials can also be employed with liquid carriers to form spreadablepastes, gels, ointments, soaps, and the like, for application directlyto the skin of the user.

The compound or a pharmaceutically acceptable prodrug or salts thereofcan also be mixed with other active materials that do not impair thedesired action, or with materials that supplement the desired action,such as antibiotics, antifungals, anti-inflammatories, or otherantibacterials, including other nucleoside compounds. Solutions orsuspensions used for parenteral, intradermal, subcutaneous, or topicalapplication can include the following components: a sterile diluent suchas water for injection, saline solution, fixed oils, polyethyleneglycols, glycerine, propylene glycol or other synthetic solvents;antibacterial agents such as benzyl alcohol or methyl parabens;antioxidants such as ascorbic acid or sodium bisulfite; chelating agentssuch as ethylenediaminetetraacetic acid; buffers such as acetates,citrates or phosphates and agents for the adjustment of tonicity such assodium chloride or dextrose. The parental preparation can be enclosed inampoules, disposale syringes or multiple dose vials made of glass orplastic. If administered intravenously, useful carriers arephysiological saline or phosphate buffered saline (PS).

In one embodiment, the active compounds are prepared with carriers thatwill protect the compound against rapid elimination from the body, suchas a controlled release formulation, including implants andmicroencapsulated delivery systems. Biodegradable, biocompatiblepolymers can be used, such as ethylene vinyl acetate, polyanhydrides,polyglycolic acid, collagen, polyorthoesters and polylactic acid.Methods for preparation of such formulations will be apparent to thoseskilled in the art. The materials can also be obtained commercially fromAlza Corporation.

The concentration of the compound(s) in a liquid composition, such as alotion, will, for example, range from about 0.1% to about 95% by weight,or from about 0.5% to about 25% by weight. The concentration in asemi-solid or solid composition such as a gel or a powder will, forexample, range from about 0.1% to 100% by weight, or about 0.5% to about5% by weight. Single doses for intravenous injection, subcutaneous,intramuscular or topical administration, infusion, ingestion orsuppository will generally be from about 0.001 to about 5000 mg, and beadministered from about 1 to about 3 times daily, to yield levels ofabout 0.01 to about 500 mg/kg, for adults.

The invention also includes one or more compounds disclosed herein, orany combination thereof, or salt thereof, in an amount effective toinhibit bacterial (such as a gram negative bacteria) replication in ahost. The compound can be useful for inhibiting bacterial replication ina cell or neutralization (i.e. inactivation) of extracellular bacteria.

As used herein, to inhibit bacterial replication in a host means toreduce the bacterial load in a host to a level which is lower than thelevel of the bacterial load in an otherwise identical host which was notadministered the compound. Bacterial load in a mammal can be reduced byabout 1 to 12 log₁₀ or more relative to an otherwise identical mammalwhich was not administered the compound. Bacterial load in a mammal canbe assessed by a number of methods known in the art such as, forexample, obtaining a tissue or fluid sample from the mammal andassessing the amount of bacterial components in the mammal containedtherein using technology which is either immunological, biochemical ormolecular biological in nature and which is well known to the skilledartisan and which are described elsewhere herein. Inhibition ofbacterial replication in a cell is assessed using similar or identicalassays as those used to assess bacterial load in a mammal.

The invention also includes a kit for administering a compound of theinvention, a pharmaceutically acceptable salt thereof, or apharmaceutical composition, to a host for treatment of a bacterial (suchas a gram negative bacteria) infection. Typically, the host is a human.The kit comprises one or more compounds of the invention, or acombination thereof, and optionally an instructional material, whichdescribes adventitially administering the composition to the mammal byany of the routes of administration described herein. In anotherembodiment, this kit comprises a (typically sterile) solvent suitablefor dissolving or suspending the composition of the invention prior toadministering the compound to the mammal.

EXAMPLES

Melting points were determined on a MeI-temp II laboratory device andare uncorrected. Nuclear magnetic resonance spectra were obtained on aGE 300 Plus (300 MHz), a Varian INOVA 400 (400 MHz), and a Varian INOVA600 (600 MHz) spectrometer; chemical shifts (δ) are reported in partsper million (ppm), and the signals are described as s (singlet), d(doublet), t (triplet), q (quartet), bs or brs (broad singlet), dd(doublet of doublet), and m (multiplet). UV spectra were obtained on aBeckman DU 650 spectrophotometer. Mass spectra were measured on aMicromass Inc. Autospec High Resolution double focussing sector (EBE) MSspectrometers. Infrared spectra were obtained on a Nicolet 510 FT-IRspectrometer. All reactions were monitored using thin layerchromatography on Analtech, 200 mm silica gel GF plates. Dry1,2-dichloroethane, dichloromethane, acetonitrile,N,N-dimethylformamide, and THF were obtained by drying over 4 Amolecular sieves.

ABBREVIATIONS

ACN: acetonitrileDCE: 1,2-dichloroethaneDCM: dichloromethaneDDQ: dichlorodicyano quinoneDIEA: diisopropylethyl amineDI H₂O: deionized waterDMAP: 4-dimethylamino pyridineDMF: N,N-dimethyl formamideDPPB: 1,4-bis(diphenylphosphine)butaneLAH: lithiumaluminum hydrideLHMDS: lithium hexamethyldisilazidePd/C: palladium on carbonPNB: p-nitrobenzylPTSA: p-toluenesulfonic acid monohydrateTBAF: tetrabutylammonium fluorideTBDMS: t-butyldimethylsilylTEA: triethylamineTES: triethylsilylTFA: trifluoroacetic acidTHF: tetrahydrofuranTLC: thin layer chromatographyTBDPS: t-butyldiphenylsilyl

Preparation of the Carbapenem Intermediate (CPI) 5

Carbapenem Intermediate (CPI) 5 was prepared according to the syntheticscheme shown in FIG. 3. In the first step of the process, benzylpropionate is reacted with isobutoxycarbonyloxy acetic acid methyl esterin a solvent at low temperature in the presence of LDA to form ketoesterA. The ketoester A is then contacted with the acetoxyazetidinone B(prepared by any number of known, synthetic routes) in a solvent, andsodium carbonate is added. The reaction ages for a period of time at atemperature such that the reaction goes substantially to completion,generating the target lactam C.

The lactam C is dissolved in a solvent, such as DMF, to which a suitablebase (such as DIEA) and TBSOTf are added, and the mixture allowed to agefor a period of time at a temperature. Following workup, thebis-TBS-ketoester D is isolated.

The crude ketoester D is dissolved in ethyl acetate in an appropriatereaction vessel. Formic acid and a catalyst, such as Pd/C, are added tothe reaction vessel, and the entire mixture is hydrogenated at anappropriate hydrogen pressure (40-50 psi) for a period of time such thatthe decarboxylation reaction proceeds to completion. The reactionmixture is filtered over a pad of Celite®, and the solvent is removedunder vacuum. Product E is isolated following purification by columnchromatography.

The bis-TBDMS ketolactam E is then de-silylated using 2 N HCl in ACN andthe product is isolated after a standard aqueous workup. The crudeproduct is dissolved in a solvent, such as DCM, and allowed to reactwith triethylsilyl chloride and imidazole for several hours (monitoredby TLC) at rt. Following aqueous workup, O-TES ketolactam F was isolatedand purified on silica gel.

N-PNB, O-TES ketolactam G is produced by reacting ketolactam F withp-nitrobenzyl oxalylchloride in a suitable solvent (DCM, for example) inthe presence of a base (DMA, for example). The mixture is allowed to agefor a period of time (and at an appropriate temperature) to effect asubstantially complete reaction as monitored by an appropriate means(e.g., TLC or HPLC). Following workup in a usual manner, intermediate Gwas isolated.

To a solution of compound G is a suitable solvent was addedtriethylphosphite, and the mixture heated to reflux until complete byTLC. Following workup and purification in the appropriate manner, CPI 5was isolated.

The Amine Series: Preparation of Gram-Negative Active Carbapenems

The amine series of 1-β-methylcarbapenem analogs possessingGram-negative activity were synthesized using synthetic methodsdescribed above and as illustrated in Scheme 2 below, unless otherwisenoted. In general, a series of secondary amines (2) were coupled to CPI5 (FIG. 3) in DMF using a combination of Pd₂(dba)₃CHCl₃ with DPPB(Method A) or triethylphosphite (Method B) at it to produce the coupledintermediate 3. In some cases, 2,6-lutidine (Method C) or PTSA (MethodD) were added to drive the reaction to completion. The secondary amineswere either purchased from commercial sources or prepared by alkylationof N-Boc-protected primary amines with various alkyl halides followed bycleavage of the Boc protecting group with TFA/water in DCM.

Removal of the TES protecting group in the series of intermediate 3 wasaccomplished with aqueous triflic acid solution in IPA/water (pH 2.4,Method E) or with 0.06N HCl in IPA/THF (Method F) at 0° C. to rt.

Lastly, the PNB group(s) in intermediate 4 were removed by hydrogenationof the corresponding PNB esters using standard conditions (atmosphericH₂ pressure, 5% Pt/C, THF/i-PrOH/0.5 M potassium phosphate buffersolution (pH 7.0) at 0° C.) and the final products 6 were isolated afterpurification on SP-207 resin.

Step 1: General Procedure for the Palladium Coupling Reaction Method A

To a dry, round-bottom flask, anhydrous DMF (40 mL) was added and thesolvent was stirred and degassed at room temperature with two cycles ofnitrogen/vacuum. Pd₂(dba)₃CHCl₃ (51.8 mg, 0.05 mmol) and DPPB (64.0 mg,0.15 mmol) were then added. The solution was degassed with twonitrogen/vacuum cycles and aged for 20 minutes. The appropriatesecondary amine (2) (1.1 mmol) was added and the mixture was againdegassed under reduced pressure for 5 min. CPI 5 (590.7 mg, 1.0 mmol)was then added, the resulting mixture was again degassed with twonitrogen/vacuum cycles, and the resulting mixture was aged for 12-36 h(monitored by TLC) at rt. The solvent was then removed under reducedpressure and the resulting residue was purified by flash chromatographyon silica gel to produce coupled intermediate 3 in 50-98% yield.

Method B

To a dry, round-bottom flask, anhydrous DMF (40 mL) was added and thesolvent was stirred and degassed at room temperature with two cycles ofnitrogen/vacuum. Pd₂(dba)₃CHCl₃ (51.8 mg, 0.05 mmol) and P(OEt)₃ (24.9mg, 0.15 mmol). were then added. The solution was degassed with twonitrogen/vacuum cycles and aged for 20 minutes. The appropriatesecondary amine (2) (1.1 mmol) was added and the mixture was againdegassed under reduced pressure for 5 min. CPI 5 (590.7 mg, 1.0 mmol)was then added, the resulting mixture was again degassed with twonitrogen/vacuum cycles, and the resulting mixture was aged for 12-36 h(monitored by TLC) at rt. The solvent was then removed under reducedpressure and the resulting residue was purified by flash chromatographyon silica gel to produce coupled intermediate 3 in 50-98% yield.

Method C

Used a similar procedure as was described in Method B except that2.6-lutidine (0.5 eq) was added in conjunction with reagent 2.

Method D

Used a similar procedure as was described in Method B except that PTSA(0.5 eq) and 4 A molecular sieves were added in conjunction with thepalladium catalyst.

Step 2: General Procedure for the Removal of the TES Protecting GroupMethod E

Preparation of the pH 2.4 aqueous/IPA triflic acid stock solution; 620uL of triflic acid was added to a stirred solution of DI H₂O (100 mL)and IPA (500 mL). Additional triflic acid was added dropwise to adjustthe pH to 2.4. TES-Intermediate 3 (100 mg) was then added to 3 mL of theaqueous triflic acid:IPA stock solution and the resulting mixture wasstirred at rt. The reaction pH was then adjusted to 2.4 by theincremental addition of triflic acid stock solution, if necessary. Thereaction mixture was stirred at rt until complete by TLC (˜2 h),neutralized with 0.5M potassium phosphate buffer solution (pH 7.0, 5-10mL), and the resulting mixture was stirred for 10 min and then extractedwith ethyl acetate. The organic layer was washed with brine, dried overanhydrous sodium sulfate, and concentrated under reduced pressure. Thecrude product was purified by flash chromatography on silica gel usingas eluent hexane/ethyl acetate with gradient or ethyl acetate/methanolwith gradient to produce alcohol 4 in 67-92% yield.

Method F

Alcohol 4 (100 mg) was dissolved in THF/IPA (2 mL/2 mL) and cooled to 0°C. 0.06N HCl solution was then added (1-3 mL, pH adjusted to 2.4) andthe resulting mixture was aged at 0° C. for 2-18 h (monitored by TLC).The reaction was neutralized with 0.5M potassium phosphate buffersolution (pH 7.0, 5-10 mL), and the resulting mixture was carried on toStep 3 without further isolation or purification.

Step 3: General Procedure for the Removal of the PNB Protecting Group

To a round-bottom flask equipped with a side arm were added alcoholintermediate 4 (200 mg) and THF/IPA (3 mL/3 mL), and the resultingmixture was stirred at rt. Aqueous 0.5 M potassium phosphate buffersolution (pH 7.0, 6 mL), was then added and the resulting mixture wascooled to 0° C. 5% Pt/C (40 mg) was added and the reaction flask wasfitted with a hydrogen balloon, degassed under reduced pressure, andcharged with hydrogen. The resulting mixture was aged for 4-14 hrs.(monitored by TLC) at 0° C., diluted with chilled ethyl acetate (20 mL)and DI H₂O (10 mL), and filtered over a pad of Celite. The celite waswashed with chilled DI H₂O (30 mL) and ethyl acetate (30 mL), the layersseparated, and the aqueous layer was extracted with cold ethyl acetate(30 mL). The aqueous fraction was then concentrated under reducedpressure to remove any organics and lyophilized. The crude material waspurified on SP-207 resin with IPA/DI H₂O and the column fractionsconcentrated under reduced pressure at 10° C. to remove i-PrOH and thenlyophilized to give the desired, final product 6 as fluffy solids.

Example 1 TES-Protected Cyanoethylamine CP Intermediate 7

Method A; Percent yield: 98%; ¹H NMR (CDCl₃, 300 MHz): δ 8.20 (d, J=9.0Hz, 2H), 7.66 (d, J=8.4 Hz, 2H), 5.44 (d, J=14.1 Hz, 2H), 5.21 (d,J=14.1 Hz, 2H), 4.29-4.20 (m, 2H), 3.93 (d, J=15.0 Hz, 1H), 3.47-3.36(m, 1H), 3.25 (dd, J=4.8; 2.7 Hz, 1H), 3.14 (d, J=14.7 Hz, 1H),2.78-2.69 (m, 1H), 2.64-2.56 (m, 1H), 2.51-2.47 (m, 2H), 2.25 (s, 3H),1.24 (d, J=7.2 Hz, 3H), 1.18 (d, J=7.5 Hz, 3H), 0.93 (t, J=7.8 Hz, 9H),0.59 (q, J=7.5 Hz, 6H).

PNB-Protected Cyanoethylamine CP Intermediate 8

Method E; Percent yield: 75%; ¹H NMR (CDCl₃, 300 MHz): δ 8.16 (d, J=9.3Hz, 2H), 7.60 (d, J=8.4 Hz, 2H), 5.43 (d, 14.4 Hz, 2H), 5.15 (d, J=14.4Hz, 2H), 4.24-4.18 (m, 2H), 3.90 (d, J=14.7 Hz, 1H), 3.50-3.40 (m, H),3.26 (dd, J=6.6; 3.0 Hz, 1H), 3.11 (d, J=14.7 Hz, 1H), 2.86 (br s, 1H),2.75-2.66 (m, 1H), 2.60-2.52 (m, 1H), 2.50-2.45 (m, 2H), 2.23 (s, 3H),1.28 (d, J=6.9 Hz, 3H), 1.15 (d, J=7.5 Hz, 3H).

Cyanoethylamine CP Analog 9

Percent yield: 50%; ¹H NMR (D₂O, 400 MHz): δ 4.22-4.15 (m, 2H), 3.75 (d,J=14.4 Hz, 1H), 3.42-3.39 (m, 2H), 3.24-3.16 (m, 1H), 3.05-2.98 (m, 1H),2.83-2.72 (m, 3H), 2.38 (s, 3H), 1.23 (d, J=6.4 Hz, 3H), 1.08 (d, J=7.6Hz, 3H).

Example 2 TES-Protected N-Methyl Acetamide CP Intermediate 10

Method A; Percent yield; 98%; ¹H NMR (CDCl₃, 300 MHz): δ 8.20 (d, J=8.1Hz, 2H), 7.65 (d, J=8.4 Hz, 2H), 6.87 (br s, 1H), 6.32 (br s, 1H), 5.43(d, J=14.1 Hz, 1H), 5.21 (d, J=14.1 Hz, 1H), 4.29-4.19 (m, 2H), 4.03 (d,J=14.7 Hz, 1H), 3.33-3.23 (m, 2H), 3.14 (14.1 Hz, 1H), 3.07 (d, J=17.1Hz, 1H), 2.93 (d, J=15.6 Hz, 1H), 2.29 (s, 3H), 1.24 (d, J=5.4 Hz, 3H),1.18 (d, J=7.2 Hz, 3H), 0.92 (t, J=8.4 Hz, 9H), 0.58 (q, J=8.4 Hz, 6H).

PNB-Protected N-Methyl Acetamide CP Intermediate 11

Method E; Percent yield: 75%; ¹H NMR (CDCl₃, 300 MHz): δ 8.16 (d, J=8.4Hz, 2H), 7.60 (d, J=8.7 Hz, 2H), 6.88 (br s, 1H), 6.50 (br s, 1H), 5.43(d, J=14.1 Hz, 2H), 5.17 (d, J=14.1 Hz, 2H), 4.23-4.15 (m, 2H), 3.97 (d,J=13.5 Hz, 1H), 3.36-3.24 (m, 2H), 3.11 (d, J=14.1 Hz, 1H), 103 (d,J=15.6 Hz, 1H), 2.88 (d, J=16.8 Hz, 1H), 2.25 (s, 3H), 1.28 (d, J=5.7Hz, 3H), 1.15 (d, J=7.2 Hz, 3H).

N-Methyl Acetamide CP Analog 12

Percent yield: 87%; ¹H NMR (D₂O, 400 MHz): δ 4.26-4.18 (m, 2H), 3.92 (d,J=14.4 Hz, 1H), 3.66 (d, J=14.4 Hz, 1H), 3.61 (d, J=16.0 Hz, 1H), 3.52(d, J=16.0 Hz, 1H), 3.46-3.44 (m, 1H), 3.29-3.21 (m, 1H), 2.59 (s, 3H),1.27 (d, J=6.4 Hz, 3H), 1.13 (d, J=7.2 Hz, 3H).

Example 3 TES-Protected N-Methyl Acetamide CP Intermediate 13

Method C; Percent yield: 62%; ¹H NMR (CDCl₃, 300 MHz): δ 8.21 (d, J=8.4Hz, 2H), 7.66 (d, J=9.3 Hz, 2H), 6.90 (br s 1H), 6.22 (br s, 1H), 5.44(d, J=13.5 Hz, 1H), 5.38 (br s, 1H), 5.22 (d, J=13.5 Hz, 1H), 4.28-4.20(m, 2H), 4.04 (d, J=14.1 Hz, 1H), 3.30-3.24 (m, 1H), 3.15 (d, J=14.1 Hz,1H), 3.08 (d, J=16.2 Hz, 1H), 2.94 (d, J=16.2 Hz, 1H), 2.30 (s, 3H),1.25 (d, J=6.6 Hz, 3H), 1.18 (d, J=6.6 Hz, 3H), 0.93 (t, J=8.4 Hz, 9H),0.59 (q, J=7.8 Hz, 6H).

PNB-Protected N-Methyl Acetamide CP Intermediate 14

Method E; Percent yield: 60%; ¹H NMR (acetone-d₆, 300 MHz): δ 8.22 (d,J=9.0 Hz, 2H), 7.79 (d, J=8.4 Hz, 2H), 7.15 (br s, 1H), 6.76 (br s, 1H),5.51 (d, J=14.1 Hz, 2H), 5.30 (d, J=14.1 Hz, 2H), 4.26 (dd, J=10.2; 3.0Hz, 1H), 4.18-4.07 (m, 1H), 4.06-3.94 (m, 2H), 3.56-3.45 (m, 1H),3.33-3.30 (m, 1H), 3.07 (d, J=15.6 Hz, 1H), 2.94 (d, J=15.6 Hz, 1H),2.28 (s, 3H), 1.24 (d, J=6.0 Hz, 3H), 1.20 (d, J=7.5 Hz, 3H).

N-Methyl Acetamide CP Analog 15

Percent yield: 46%; ¹H NMR (D₂O, 300 MHz): δ 4.22-4.13 (m, 2H), 3.86 (d,J=13.5 Hz, 1H), 3.62-3.41 (m, 3H), 3.40 (dd, J=6.0; 2.7 Hz, 1H),3.26-3.16 (m, 1H), 2.52 (s, 3H), 1.22 (d, J=6.0 Hz, 1H), 1.09 (d, J=7.5Hz, 3H).

Example 4 TES-Protected N-Methyl Ethylamine CP Intermediate 16

Method A; Percent yield: 48%; ¹H NMR (CDCl₃, 300 MHz): δ 8.18 (d, J=8.4Hz, 2H), 8.17 (d, J=8.7 Hz, 2H), 7.64 (d, J=8.4 Hz, 2H), 7.48 (d, J=8.4Hz, 2H), 5.43 (d, J=13.8 Hz, 1H), 5.29 (br s, 1H), 5.19 (d, J=13.8 Hz,1H), 5.16 (s, 2H), 4.25-4.16 (m, 2H), 3.85 (d, J=14.4 Hz, 1H), 3.32-3.26(m, 3H), 3.22 (dd, J=5.7; 2.7 Hz, 1H), 3.10 (d, J=14.4 Hz, 1H),2.56-2.42 (m, 2H), 2.18 (s, 3H), 1.22 (d, J=6.6 Hz, 3H), 1.13 (d, J=7.2Hz, 3H), 0.90 (t, J=8.4 Hz, 9H), 0.57 (q, J=7.8 Hz, 6H).

PNB-Protected N-Methyl Ethylamine CP Intermediate 17

Method E; Percent yield: 93%; ¹H NMR (CDCl₃, 300 MHz): δ 8.19 (d, J=8.4Hz, 2H), 8.18 (d, J=7.5 Hz, 2H), 7.62 (d, J=8.4 Hz, 2H), 7.49 (d, J=8.4Hz, 2H), 5.46 (d, J=13.8 Hz, 1H), 5.40 (br s, 1H), 5.18 (d, J=13.8 Hz,1H), 5.17 (s, 2H), 4.23 (m, 2H), 3.90 (d, J=15.0 Hz, 1H), 3.38-3.25 (m,4H), 3.16 (d, J=14.4 Hz, 1H), 2.64-2.44 (m, 2H), 2.23 (s, 3H), 1.30 (d,J=5.4 Hz, 3H), 1.14 (d, J=7.5 Hz, 3H).

N-Methyl Ethylamine CP Analog 18

Percent yield: 27%; ¹H NMR (D₂O, 300 MHz): δ 4.20-4.13 (m, 2H), 3.76 (d,J=14.7 Hz, 1H), 3.40-3.32 (m, 2H), 3.19-3.10 (m, 3H), 2.91-2.68 (m, 2H),2.36 (s, 3H), 1.22 (d, J=6.0 Hz, 3H), 1.02 (d, J=7.2 Hz, 3H).

Example 5 TES-Protected N-Methyl Sulfonamide CP Intermediate 19

Method C; Percent yield: 43%; ¹H NMR (CDCl₃, 300 MHz): δ 8.22 (d, J=8.4Hz, 2H), 7.68 (d, J=9.0 Hz, 2H), 5.46 (d, J=14.1 Hz, 1H), 5.36 (br s,1H), 5.24 (d, J=14.1 Hz, 1H), 5.10 (br s, 1H), 4.29-4.22 (m, 2H), 3.96(d, J=13.8 Hz, 1H), 3.31-3.21 (m, 3H), 3.08 (d, J=13.8 Hz, 1H),2.70-2.50 (m, 2H), 2.24 (s, 3H), 1.25 (d, J=6.3 Hz, 3H), 1.16 (d, J=7.5Hz, 3H), 0.95 (t, J=8.7 Hz, 9H), 0.60 (q, J=7.5 Hz, 6H).

PNB-Protected N-Methyl Sulfonamide CP Intermediate 20

Method E; Percent yield: 61%; ¹H NMR (acetone-d₆, 300 MHz): δ 8.23 (d,J=9.0 Hz, 2H), 7.79 (d, J=9.0 Hz, 2H), 5.98 (br s, 1H), 5.74 (d, J=14.1Hz, 1H), 5.45 (br s, 1H), 5.29 (d, J=14.1 Hz, 1H), 4.24 (dd, J=10.2; 3.6Hz, 1H), 4.20-4.09 (m, 1H), 3.84 (d, J=14.7 Hz, 1H), 3.48-3.40 (m, 1H),3.30 (dd, J=7.2; 3.0 Hz, 1H), 3.25-3.15 (m, 1H), 2.67-2.58 (m, 1H),2.51-2.45 (m, 1H), 2.21 (s, 3H), 1.24 (d, J=6.3 Hz, 3H), 1.18 (d, J=6.6Hz, 3H).

N-Methyl Sulfonamide CP Analog 21

Percent yield: 68%; ¹H NMR (D₂O, 400 MHz): δ 4.25-4.22 (m, 2H), 4.06 (d,J=14.4 Hz, 1H), 3.95 (d, J=14.4 Hz, 1H), 3.49 (dd, J=6.0; 2.8 Hz, 1H),3.48-3.45 (m, 2H), 3.34-3.19 (m, 3H), 2.85 (s, 3H), 1.27 (d, J=6.4 Hz,3H), 1.17 (d, J=7.6 Hz, 3H).

Example 6 TES-Protected N-Methyl Imidazole CP Intermediate 22

Method C; Percent yield: 40%; ¹H NMR (CDCl₃, 300 MHz): δ 8.15 (d, J=9.0Hz, 2H), 7.62 (d, J=8.4 Hz, 2H), 7.44 (s, 1H), 6.97 (s, 1H), 6.88 (s,1H), 5.40 (d, J=13.8 Hz, 1H), 5.16 (d, J=13.8 Hz, 1H), 4.23-4.16 (m,1H), 4.05 (dd, J=10.5; 3.3 Hz, 1H), 3.98-3.95 (m, 2H), 3.89 (d, J=15.0Hz, 1H), 3.14 (dd, J=4.8; 3.0 Hz, 1H), 3.02 (d, J=15.0 Hz, 1H),2.88-2.77 (m, 1H), 2.71-2.62 (m, 1H), 2.59-2.50 (m, 1H), 2.23 (s, 3H),1.18 (d, J=6.6 Hz, 3H), 0.96-0.88 (m, 12H), 0.55 (q, J=8.1 Hz, 6H).

N-Methyl Imidazole CP Analog 23

Method F; Percent yield: 38% (from 22); ¹H NMR (D₂O, 300 MHz): δ 7.79(s, 1H), 7.16 (s, 1H), 7.03 (s, 1H), 4.32-4.25 (m, 2H), 4.20-4.11 (m,1H), 4.08-4.05 (m, 1H), 3.73 (d, J=15.0 Hz, 1H), 3.42 (d, J=15.0 Hz,1H), 3.36-3.40 (m, 1H), 3.19-3.10 (m, 1H), 2.99-2.86 (m, 2H), 2.46 (s,3H), 1.20 (d, J=6.3 Hz, 3H), 1.00 (d, J=7.2 Hz, 3H).

Example 7 TES-Protected N-Methyl Guanidine CP Intermediate 24

Method B; Percent yield: 32%; ¹H NMR (CDCl₃, 300 MHz): δ 11.77 (s, 1H),8.68 (s, 1H), 8.26-8.19 (m, 6H), 7.67 (d, J=8.1 Hz, 2H), 7.54-7.50 (m,4H), 5.46 (d, J=13.5 Hz, 1H), 5.25 (s, 2H), 5.22 (s, 2H), 5.22 (d,J=13.5 Hz, 1H), 4.28-4.20 (m, 2H), 3.93 (d, J=14.7 Hz, 1H), 3.57-3.55(m, 2H), 3.46-3.38 (m, 1H), 3.24 (dd, J=5.4; 3.0 Hz, 1H), 3.16 (d,J=14.7 Hz, 1H), 2.68-2.48 (m, 2H), 2.21 (s, 3H), 1.25 (d, J=6.0 Hz, 3H),1.13 (d, J=7.2 Hz, 3H), 0.93 (t, J=8.1 Hz, 9H), 0.60 (q, J=8.1 Hz, 6H).

N-Methyl Guanidine CP Analog 25

Method F; Percent yield: 25% (from 24); ¹H NMR (D₂O, 400 MHz): δ 4.23(quintet, J=6.0 Hz, 1H), 4.18 (d, J=10.0 Hz, 1H), 3.75 (d, J=14.4 Hz,1H), 3.44-3.38 (m, 4H), 3.18 (quintet, J=8.4 Hz, 1H), 2.90-2.80 (m, 1H),2.70-2.60 (m, 1H), 2.38 (s, 3H), 1.27 (d, J=6.4 Hz, 3H), 1.11 (d, J=7.2Hz, 3H).

Example 8 TES-Protected N-Methyl Thiazole CP Intermediate 26

Method A; Percent yield: 77%; ¹H NMR (CDCl₃, 300 MHz): δ 8.22 (d, J=7.8Hz, 2H), 7.71 (d, J=4.2 Hz, 1H), 7.67 (d, J=9.0 Hz, 2H), 7.30 (d, J=3.0Hz, 1H), 5.46 (d, J=13.8 Hz, 1H), 5.23 (d, J=13.8 Hz, 1H), 4.27-4.22 (m,1H), 4.19 (dd, J=10.2; 3.0 Hz, 1H), 4.02 (d, J=14.7 Hz, 1H), 3.93 (d,J=15.0 Hz, 1H), 3.80 (d, J=15.0 Hz, 1H), 3.46-3.38 (m, 1H), 3.29-3.23(m, 2H), 2.33 (s, 3H), 1.26 (d, J=5.7 Hz, 3H), 1.17 (d, J=7.2 Hz, 3H),0.94 (t, J=8.4 Hz, 9H), 0.60 (q, J=7.8 Hz, 6H).

PNB-Protected N-Methyl Thiazole CP Intermediate 27

Method E; Percent yield: 98%; ¹H NMR (CDCl₃, 300 MHz): δ 8.14 (d, J=6.3Hz, 2H), 7.64 (d, J=3.6 Hz, 1H), 7.59 (d, J=8.7 Hz, 2H), 7.25 (d, J=3.0Hz, 1H), 5.43 (d, J=13.5 Hz, 1H), 5.16 (d, J=13.5 Hz, 1H), 4.20 (dd,J=9.6; 3.0 Hz, 2H), 3.96 (d, J=14.7 Hz, 1H), 3.87 (d, J=15.3 Hz, 1H),3.75 (d, J=15.3 Hz, 1H), 3.48-3.30 (m, 2H), 3.26-3.19 (m, 2H), 2.26 (s,3H), 1.28 (d, J=6.0 Hz, 3H), 1.12 (d, J=7.5 Hz, 3H).

N-Methyl Thiazole CP Analog 28

Percent yield: 54%; ¹H NMR (D₂O, 400 MHz): δ 7.76 (d, J=3.6 Hz, 1H),7.60 (d, J=3.6 Hz, 1H), 4.22 (d, J=14.0 Hz, 1H), 4.17 (quint, J=6.4 Hz,1H), 4.11 (dd, J=10.0; 3.2 Hz, 1H), 4.04 (d, J=14.4 Hz, 1H), 3.80 (d,J=14.4 Hz, 1H), 3.45 (d, J=14.4 Hz, 1H), 3.36 (dd, J=6.4; 2.8 Hz, 1H),3.20-3.15 (m, 1H), 2.44 (s, 3H), 1.22 (d, J=6.4 Hz, 3H), 1.01 (d, J=7.6Hz, 3H).

Example 9 TES-Protected Pyrrolidine CP Intermediate 29

Method D; Percent yield: 69%; ¹H NMR (CDCl₃, 300 MHz): δ 8.16 (d, J=8.1Hz, 2H), 7.63 (d, J=9.3 Hz, 2H), 5.41 (d, J=14.1 Hz, 1H), 5.18 (d,J=14.1 Hz, 1H), 4.29-4.19 (m, 2H), 4.16 (dd, J=10.2; 2.4 Hz, 1H), 3.84(d, J=14.1 Hz, 1H), 3.36-3.29 (m, 2H), 3.20 (dd, J=4.8 Hz; 2.7 Hz, 1H),2.85-2.78 (m, 1H), 2.73 (br s, 1H), 2.60 (d, J=2.7 Hz, 2H), 2.28-2.08(m, 2H), 1.72-1.66 (m, 1H), 1.22 (d, J=5.7 Hz, 3H), 1.13 (d, J=7.5 Hz,3H), 0.90 (t, J=7.2 Hz, 9H), 056 (q, J=7.8 Hz, 6H).

Pyrrolidine CP Analog 30

Method F; Percent yield: 60% (from 29); ¹H NMR (D₂O, 300 MHz): δ4.67-4.64 (m, 1H), 4.22-4.14 (m, 2H), 4.07 (d, J=14.4 Hz, 1H), 3.94 (d,J=14.4 Hz, 1H), 3.50 (br s, 1H), 3.42 (dd, J=6.3; 3.0 Hz, 1H), 3.35-3.10(m, 4H), 2.31-2.21 (m, 1H), 2.02-1.93 (m, 1H), 1.22 (d, J=6.3 Hz, 3H),1.12 (d, J=7.5 Hz, 3H).

Example 10 TES-Protected Prolineamide CP Intermediate 31

Method D; Percent yield: 85%; ¹H NMR (CDCl₃, 300 MHz): δ 8.16 (d, J=8.4Hz, 2H), 7.62 (d, J=8.7 Hz, 2H), 7.02 (d, J=4.5 Hz, 1H), 6.68 (d, J=4.5Hz, 1H), 5.40 (d, J=14.1 Hz, 1H), 5.19 (d, J=14.1 Hz, 1H), 4.24-4.17 (m,2H), 4.06 (d, J=14.4 Hz, 1H), 3.35-3.20 (m, 3H), 3.06 (dd, J=9.3; 4.8Hz, 1H), 3.00-2.96 (m, 1H), 2.29-2.12 (m, 2H), 1.91-1.81 (m, 1H),1.78-1.67 (m, 2H), 1.21 (d, J=6.6 Hz, 3H), 1.12 (d, J=7.5 Hz, 3H), 0.89(t, J=8.4 Hz, 9H), 055 (q, J=8.4 Hz, 6H).

Prolineamide CP Analog 32

Method F; Percent yield: 26% (from 31); ¹H NMR (D₂O, 300 MHz): δ4.22-4.12 (m, 2H), 3.89 (d, J=13.8 Hz, 1H), 3.65-3.53 (m, 2H), 3.42-3.20(m, 3H), 2.68-2.53 (m, 1H), 2.40-2.23 (m, 1H), 1.98-1.75 (m, 3), 1.22(d, J=5.7 Hz, 3H), 1.06 (d, J=7.5 Hz, 3H).

Example 11 TES-Protected Piperazine CP Intermediate 33

Method D; Percent yield: 37%; ¹H NMR (CDCl₃, 300 MHz): δ 8.20 (d, J=8.1Hz, 2H), 7.66 (d, J=9.3 Hz, 1H), 5.43 (d, J=14.1 Hz, 1H), 5.22 (d,J=14.1 Hz, 1H), 4.97 (br s, 1H), 4.28-4.20 (m, 2H), 3.79 (d, J=14.1 Hz,1H), 3.34-2.94 (m, 7H), 2.89-2.82 (m, 1H), 2.65-2.60 (m, 1H), 2.26 (t,J=9.9 Hz, 1H), 1.85 (t, J=10.2 Hz, 1H), 1.24 (d, J=6.3 Hz, 3H), 1.15 (d,J=7.8 Hz, 3H), 0.93 (t, J=8.4 Hz, 9H), 0.59 (q, J=8.1 Hz, 6H).

Piperazine CP Analog 34

Method F; Percent yield: 58% (from 33); ¹H NMR (D₂O, 300 MHz): δ4.20-4.10 (m, 2H), 3.70 (d, J=13.8 Hz, 1H), 3.45-3.36 (m, 2H), 3.23-2.89(m, 8H), 2.53 (t, J=10.8 Hz, 1H), 2.10 (t, J=10.5 Hz, 1H), 1.20 (d,J=6.0 Hz, 3H), 1.05 (d, J=7.5 Hz, 3H).

Example 12 TES-Protected Piperazine CP Intermediate 35

Method D; Percent yield: 13%; ¹H NMR (CDCl₃, 300 MHz): δ 8.19 (d, J=8.4Hz, 2H), 7.65 (d, J=8.1 Hz, 2H), 6.40 (br s, 4H), 5.40 (d, J=14.1 Hz,1H), 5.21 (d, J=14.1 Hz, 1H), 4.26-4.18 (m, 2H), 3.37 (d, J=14.1 Hz,1H), 3.49-3.39 (m, 3H), 3.26-3.00 (m, 5H), 2.94-2.90 (m, 1H), 2.78-2.74(m, 1H), 2.52-2.42 (m, 1H), 1.22 (d, J=6.9 Hz, 3H), 1.15 (d, J=6.0 Hz,3H), 0.92 (t, J=8.4 Hz, 9H), 0.57 (q, J=7.8 Hz, 6H).

Piperazine CP Analog 36

Method F; Percent yield: 63% (from 35); ¹H NMR (D₂O, 300 MHz): δ4.18-4.10 (m, 2H), 3.72 (d, J=14.4 Hz, 1H), 3.43-3.35 (m, 2H), 3.21-2.88(m, 8H), 2.36-2.27 (m, 2H), 1.20 (d, J=6.0 Hz, 3H), 1.05 (d, J=7.2 Hz,3H).

Example 13 TES-Protected Prolineamide CP Intermediate 37

Method D; Percent yield: 91%; ¹H NMR (CDCl₃, 300 MHz): δ 8.23 (d, J=9.0Hz, 2H), 7.67 (d, J=9.0 Hz, 2H), 7.06 (br s, 1H), 5.46 (d, J=13.8 Hz,1H), 5.24 (d, J=13.8 Hz, 1H), 5.43 (br s, 1H), 4.30-4.22 (m, 2H),4.17-4.07 (m, 2H), 3.38-3.24 (m, 2H), 3.14 (dd, J=9.9; 6.0 Hz, 1H),3.05-3.01 (m, 1H), 2.36-2.19 (m, 2H), 1.98-1.72 (m, 3H), 1.26 (d, J=6.3Hz, 3H), 1.17 (d, J=7.5 Hz, 3H), 0.95 (t, J=8.1 Hz, 9H), 0.61 (q, J=8.1Hz, 6H).

PNB-Protected Prolineamide CP Intermediate 38

Method E; Percent yield: 96%; NMR (acetone-d₆, 300 MHz): δ 8.23 (d,J=9.0 Hz, 2H), 7.80 (d, J=9.0 Hz, 2H), 7.19 (br s, 1H), 6.45 (br s, 1H0,5.52 (d, J=14.1 Hz, 1H), 5.31 (d, J=14.1 Hz, 1H), 4.25 (dd, J=9.9; 3.6Hz, 1H), 4.06-3.99 (m, 2H), 3.62-3.52 (m, 1H), 3.37-3.29 (m, 2H),3.06-2.87 (m, 3H), 2.28-2.10 (m, 2H), 1.86-1.72 (m, 3H), 1.24 (d, J=7.2Hz, 3H), 1.18 (d, J=7.2 Hz, 3H).

Prolineamide CP Analog 39

Percent yield: 62%; ¹H NMR (D₂O, 300 MHz): δ 4.24-4.16 (m, 2H), 4.04 (d,J=14.1 Hz, 1H), 3.92-3.80 (m, 2H), 3.53-3.50 (m, 1H), 3.44-3.41 (m, 1H),3.30-3.20 (m, 1H), 2.91-2.82 (m, 1H), 2.50-2.40 (m, 1H), 2.07-1.93 (m,3H), 1.24 (d, J=6.0 Hz, 3H), 1.10 (d, J=6.9 Hz, 3H).

Example 14 TES-Protected N-Methyl-2-Pyridylethyl CP Intermediate 40

Method A; Percent yield: 84%; ¹H NMR (CDCl₃, 300 MHz): δ 8.48 (d, J=3.9Hz, 1H), 8.17 (d, J=8.7 Hz, 2H), 7.63 (d, J=9.3 Hz, 2H), 7.55 (dt,J=7.8; 1.5 Hz, 1H), 7.13-7.06 (m, 2H), 5.42 (d, J=13.8 Hz, 1H), 5.19 (d,J=13.8 Hz, 1H), 4.20 (quintet, J=6.0 Hz, 1H), 4.07 (dd, J=10.5; 3.0 Hz,1H), 3.85 (d, J=14.7 Hz, 1H), 3.18-100 (m, 3H), 2.94-2.76 (3H),2.68-2.60 (m, 1H), 2.24 (s, 3H), 1.22 (d, J=6.6 Hz, 3H), 0.99 (d, J=6.6Hz, 3H), 0.92 (t, J=8.4 Hz, 9H), 0.57 (q, J=8.4 Hz, 6H).

PNB-Protected N-Methyl-2-Pyridylethyl CP Intermediate 41

Method E; Percent yield: 82%; ¹H NMR (CDCl₃, 300 MHz): δ 8.49 (d, J=5.1Hz, 1H), 8.18 (d, J=9.0 Hz, 2H), 7.63 (d, J=8.4 Hz, 2H), 7.16-7.09 (m,2H), 5.46 (d, J=13.8 Hz, 2H), 5.19 (d, J=13.8 Hz, 2H), 4.23 (m, 1H),4.15 (dd, J=9.1; 3.0 Hz, 1H), 3.87 (d, J=15.3 Hz, 1H), 3.60 (br s, 1H),3.22 (dd, J=6.0; 3.0 Hz, 1H), 3.18-3.10 (m, 2H), 2.96-2.77 (m, 3H),2.69-2.60 (m, 1H), 2.26 (s, 3H), 1.31 (d, J=6.3 Hz, 3H), 1.02 (d, J=6.3Hz, 3H).

N-Methyl-2-Pyridylethyl CP Analog 42

Percent yield: 89%; ¹H NMR (D₂O, 400 MHz): δ 8.45 (dd, J=5.2; 0.8 Hz,1H), 7.82 (dt, J=7.6; 2.0 Hz, 1H), 7.38 (d, J=8.0 Hz, 1H), 4.26-4.20 (m,2H), 4.00 (d, J=14.8 Hz, 1H), 3.93 (d, J=14.8 Hz, 1H), 3.61-3.53 (m,1H), 3.52-3.46 (m, 2H), 3.24 (t, J=6.8 Hz, 2H), 3.21-3.14 (m, 1H), 2.92(s, 3H), 1.28 (d, J=6.4 Hz, 3H), 1.18 (d, J=7.2 Hz, 3H).

Example 15 TES-Protected N-Methyl-2-Pyridylmethyl CP Intermediate 43

Method A; Percent yield: 63%; ¹H NMR (CDCl₃, 300 MHz): δ 8.55 (d, J=3.9Hz, 1H), 8.22 (d, J=8.7 Hz, 2H), 7.68-7.62 (m, 3H), 738 (d, J=8.4 Hz,1H), 7.18 (dd, J=6.6; 5.1 Hz, 1H), 5.46 (d, J=13.8 Hz, 1H), 5.23 (d,J=13.8 Hz, 1H), 4.23 (quintet, J=6.0 Hz, 1H), 4.14 (dd, J=10.2; 2.7 Hz,1H), 3.92 (d, J=14.7 Hz, 1H), 3.78 (d, J=13.2 Hz, 1H), 3.53 (d, J=13.2Hz, 1H), 3.48-3.39 (m, 1H), 3.22-3.16 (m, 1H), 2.28 (s, 3H), 1.25 (d,J=5.7 Hz, 3H), 1.07 (d, J=7.2 Hz, 3H), 0.94 (t, J=7.8 Hz, 9H), 0.59 (q,J=8.1 Hz, 6H).

PNB-Protected N-Methyl-2-Pyridylmethyl CP Intermediate 44

Method E; Percent yield: 70%; ¹H NMR (CDCl₃, 300 MHz): δ 8.55 (d, J=4.5Hz, 1H), 8.22 (d, J=8.7 Hz, 2H), 7.66 (d, J=8.4 Hz, 2H), 7.38 (d, J=7.2Hz, 2H), 7.18 (dd, J=6.6; 6.0 Hz, 1H), 5.50 (d, J=14.1 Hz, 1H), 5.22 (d,J=14.1 Hz, 1H), 4.26 (quintet, J=7.2 Hz, 1H), 4.19 (dd, J=9.9; 3.0 Hz,1H), 3.94 (d, J=14.4 Hz, 1H), 3.79 (d, J=13.2 Hz, 1H), 3.54 (d, J=13.2Hz, 1H), 3.52-3.42 (m, 2H), 3.27-3.19 (m, 2H), 2.28 (s, 3H), 1.34 (d,J=6.3 Hz, 3H), 1.09 (d, J=7.2 Hz, 3H).

N-Methyl-2-Pyridylmethyl CP Analog 45

Percent yield: 76%; ¹H NMR (D₂O, 400 MHz): δ 8.58 (d, J=4.8 Hz, 1H),7.91 (dt, J=7.6; 1.6 Hz, 1H), 7.52 (d, J=8.0 Hz, 1H), 7.49-7.46 (m, 1H),4.36 (d, J=13.6 Hz, 1H), 4.26 (d, J=13.2 Hz, 1H), 4.21-4.17 (m, 1H),4.09-4.03 (m, 2H), 3.97 (d, J=14.4 Hz, 1H), 3.42-3.40 (m, 1H), 3.19-3.11(m, 1H), 2.76 (s, 1H), 1.24 (d, J=6.4 Hz, 3H), 1.10 (d, J=7.6 Hz, 3H).

Example 16 TES-Protected N-Methyl Pyrazine CP Intermediate 46

Method A; Percent yield: 87%; ¹H NMR (CDCl₃, 300 MHz): δ 8.45 (s, 1H),8.32 (s, 1H), 8.14 (d, J=8.1 Hz, 2H), 7.61 (d, J=9.0 Hz, 2H), 5.40 (d,J=13.8 Hz, 1H), 5.16 (d, J=13.8 Hz, 1H), 4.18 (quintet, J=6.0 Hz, 1H),4.10 (dd, J=5.1; 3.0 Hz, 1H), 3.88 (d, J=14.7 Hz, 1H), 3.69 (d, J=13.8Hz, 1H), 3.48 (d, J=13.2 Hz, 1H), 3.40-3.29 (m, 1H), 3.18-3.10 (m, 2H),2.47 (s, 3H), 2.21 (s, 3H), 1.18 (d, J=5.1 Hz, 3H), 1.03 (d, J=7.2 Hz,3H), 0.87 (t, J=8.1 Hz, 9H), 0.53 (q, J=8.4 Hz, 6H).

PNB-Protected N-Methyl Pyrazine CP Intermediate 47

Method E; Percent yield: 92%; ¹H NMR (CDCl₃, 300 MHz): δ 8.47 (s, 1H),8.35 (s, 1H), 8.15 (d, J=9.0 Hz, 2H), 7.60 (d, J=9.0 Hz, 2H), 5.44 (d,J=13.8 Hz, 1H), 5.17 (d, J=13.8 Hz, 1H), 4.23-4.15 (m, 2H), 3.89 (d,J=15.0 Hz, 1H), 3.71 (d, J=13.8 Hz, 1H), 3.50 (d, J=15.0 Hz, 1H),3.44-3.36 (m, 1H), 3.23 (dd, J=6.0; 2.7 Hz, 1H), 3.15 (d, J=15.0 Hz,1H), 2.50 (s, 3H), 2.22 (s.3H), 1.27 (d, J=5.4 Hz, 3H), 1.05 (d, J=6.9Hz, 3H).

N-Methyl Pyrazine CP Analog 48

Percent yield: 63%; ¹H NMR (D₂O, 400 MHz): δ 8.52 (s, 1H), 8.47 (s, 1H),4.19-4.13 (m, 2H), 4.06-4.00 (m, 2H), 3.93 (d, J=14.4 Hz, 1H), 3.69 (d,J=14.4 Hz, 1H), 3.37-3.35 (m, 1H), 3.16-3.08 (m, 1H), 2.57 (s, 3H), 2.52(s, 3H), 1.21 (d, J=6.4 Hz, 3H), 1.03 (d, J=7.2 Hz, 3H).

Example 17 TES-Protected N-Ethyl-4-Pyridylmethyl CP Intermediate 49

Method A; Percent yield: 92%; ¹H NMR (CDCl₃, 300 MHz): δ 8.54 (d, J=5.7Hz, 2H), 8.22 (d, J=8.1 Hz, 2H), 7.67 (d, 8.4 Hz, 2H), 7.26 (d, J=5.1Hz, 2H), 5.46 (d, J=13.8 Hz, 1H), 5.23 (d, J=13.8 Hz, 1H), 4.24(quintet, J=6.0 Hz, 1H), 4.16-4.06 (m, 2H), 3.96 (d, J=15.3 Hz, 1H),3.72 (d, J=14.4 Hz, 1H), 3.37-3.14 (m, 4H), 2.62-2.55 (m, 1H), 2.42-2.33(m, 1H), 2.04 (s, 3H), 1.24 (d, J=6.3 Hz, 3H), 1.08-1.05 (m, 6H), 0.93(t, J=8.1 Hz, 9H), ) 59 (q, J=8.4 Hz, 6H).

PNB-Protected N-Ethyl-4-Pyridylmethyl CP Intermediate 50

Method E; Percent yield: 91%; ¹H NMR (CDCl₃, 300 MHz): δ 8.55 (d, J=6.3Hz, 2H), 8.23 (d, J=9.0 Hz, 2H), 7.67 (d, J=9.0 Hz, 2H), 7.29 (d, J=5.7Hz, 2H), 5.50 (d, J=13.2 Hz, 1H), 5.27 (d, J=13.2 Hz, 1H), 4.26(quintet, J=6.9 Hz, 1H), 4.18-4.11 (m, 1H), 3.97 (d, J=14.4 Hz, 1H),3.73 (d, J=14.4 Hz, 1H), 3.42-3.31 (m, 2H), 3.26 (dd, J=7.2; 3.3 Hz,1H), 3.18 (d, J=14.4 Hz, 1H), 3.65-3.54 (m, 1H), 2.45-2.35 (m, 1H), 2.30(br s, 1H), 1.35 (d, J=7.5 Hz, 3H), 1.10-1.05 (m, 6H).

N-Ethyl-4-Pyridylmethyl CP Analog 51

Percent yield: 81%; ¹H NMR (D₂O, 400 MHz): δ 8.54 (d, J=4.8 Hz, 2H),7.45 (d, J=4.8 Hz, 2H), 4.26 (d, J=13.6 Hz, 1H), 4.19-4.13 (m, 1H), 4.09(d, J=13.6 Hz, 1H), 3.99 (d, J=9.2 Hz, 1H), 3.94-3.85 (m, 2H), 3.37-3.35(m, 1H), 3.15-2.97 (m, 3H), 1.24 (t, J=7.2 Hz, 3H), 1.21 (d, J=6.4 Hz,3H), 1.05 (d, J=7.2 Hz, 3H).

Example 18 TES-Protected Pyridylthiazole CP Intermediate 52

Method A; Percent yield: 83%; ¹H NMR (CDCl₃, 300 MHz): δ 8.57 (s, 1H),8.52 (dd, J=4.8; 1.2 Hz, 1H), 8.22 (d, J=8.7 Hz, 2H), 7.69-7.61 (m, 3H),7.26 (m 1H), 5.47 (d, J=14.1 Hz, 1H), 5.23 (d, J=14.1 Hz, 1H), 4.25(quintet, J=5.7 Hz, 1H), 4.17 (dd, J=10.2; 2.7 Hz, 1H), 3.86 (d, J=14.4Hz, 1H), 3.63 (d, J=13.5 Hz, 1H), 3.43-3.31 (m, 2H), 3.23 (dd, J=5.1;3.3 Hz, 1H), 3.16 (d, J=14.4 Hz, 1H), 2.19 (s, 3H), 1.25 (d, J=6.3 Hz,3H), 1.07 (d, J=7.5 Hz, 3H), 0.94 (t, J=7.5 Hz, 9H), 0.60 (q, J=8.4 Hz,6H).

PNB-Protected Pyridylthiazole CP Intermediate 53

Method E; Percent yield: 69%; ¹H NMR (CDCl₃, 300 MHz): δ 8.74 (d, J=2.1Hz, 1H), 8.46 (d, J=5.1 Hz, 1H), 8.11 (d, J=8.4 Hz, 2H), 7.63-7.57 (m,3H), 7.42 (d, J=8.7 Hz, 1H), 7.19 (d, J=1.8 Hz, 1H), 7.11 (dd, J=5.4;4.8 Hz, 1H), 5.40 (d, J=13.8 Hz, 1H), 5.14 (d, J=13.8 Hz, 1H), 4.18-3.97(m, 3H), 3.86 (d, J=14.4 Hz, 2H), 3.72 (br s, 1H), 3.66 (d, J=13.8 Hz,1H), 3.56 (d, J=14.4 Hz, 1H), 3.48-3.37 (m, 1H), 3.20-3.13 (m, 2H), 1.23(d, J=6.3 Hz, 3H), 0.82 (d, J=7.5 Hz, 3H).

Pyridylthiazole CP Analog 54

Percent yield: 32%; ¹H NMR (D₂O, 400 MHz): δ 8.90 (s, 1H), 8.42 (d,J=4.4 Hz, 1H), 7.78 (t, J=8.0 Hz, 1H), 7.49 (s, 1H), 7.43 (d, J=7.6 Hz,1H), 7.32 (t, J=6.0 Hz, 1H), 4.10-3.86 (m, 6H), 3.77 (d, J=9.6 Hz, 1H),3.39 (d, J=14.8 Hz, 1H), 3.21-3.19 (m, 1H), 3.06 (quin, J=8.0 Hz, 1H),1.16 (d, J=5.7 Hz, 3H), 0.73 (d, J=5.4 Hz, 3H).

Preparation of Thioguanidine Analogs 62 and 63 Discussion

The following two examples of thioguanidine-substituted CP analogs wereprepared via the procedure described in Scheme 3. Alcohol intermediates55 and 58 were isolated and converted to their corresponding mesylates,56 and 59, under standard conditions. The mesylates were then allowed toreact with thiourea in dry DMF to afford the O-TES, N-PNB-protectedthioguanidine intermediates 57 and 60. Deprotection was accomplished byfollowing the two step protocol previously described.

Step I: General Procedure for Mesylate Formation

Methanesulfonyl chloride (1.3 mmol) and triethylamine (1.5 mmol) wereadded at 0° C. under nitrogen atmosphere to a solution of alcoholintermediate 55 or 58 (1.0 mmol) in dry DCM (15 mL). The reactionmixture was stirred at 0° C. for 1 h and then at 5° C. for 15 minutes(monitoring by TLC). The reaction mixture was then washed with aq.NaHCO₃, water, brine, and dried over Na₂SO₄. Evaporation of the solventgave crude mesylate, which was immediately used without purification.

Step II: General Procedure for Mesylate Displacement by Amine Sidechains

To a solution of freshly prepared mesylate (1.0 mmol) in dry DMF (15 mL)at 0° C. under N₂ atmosphere was added a solution of thiourea (1.5 mmol)in dry DMF (3 mL) and the resulting mixture was stirred at 0° C. for 1 hand then allowed to warm to rt and stirred overnight (monitored by TLC).The reaction mixture was then evaporated under reduced pressure andresidue was purified by flash chromatography. The last two deprotectionsteps were performed as previously described.

Example 19 TES-Protected N-Methyl Hydroxyethyl CP Intermediate 55

Method A; Percent yield: 74%; ¹H NMR (CDCl₃, 300 MHz): δ 8.19 (d, J=8.4Hz, 2H), 7.65 (d, J=8.4 Hz, 2H), 5.44 (d, J=14.1 Hz, 1H), 5.20 (d,J=14.1 Hz, 1H), 4.25-4.18 (m, 2H), 3.93 (d, J=14.7 Hz, 1H), 3.63-3.56(m, 2H), 3.36-3.30 (m, 1H), 3.22 (dd, J=5.7; 3.3 Hz, 1H), 3.14 (d,J=14.7 Hz, 1H), 2.70-2.56 (m, 2H), 2.49-2.41 (m, 1H), 2.21 (s, 3H), 1.22(d, J=6.3 Hz, 3H), 1.14 (d, J=7.5 Hz, 3H), 0.91 (t, J=7.5 Hz, 9H), 0.57(q, J=7.5 Hz, 6H).

TES-Protected N-Methyl Thioguanidine CP Intermediate 57

Percent yield: 54%; ¹H NMR (CDCl₃, 300 MHz): δ 9.21 (br s, 3H), 8.18 (d,J=7.8 Hz, 2H), 7.65 (d, J=8.4 Hz, 2H), 5.42 (d, J=14.1 Hz, 1H), 5.21 (d,J=14.1 Hz, 1H), 4.28-4.23 (m, 2H), 4.06 (d, J=14.7 Hz, 1H), 3.26-3.11(m, 5H), 2.82-2.73 (m, 2H), 2.26 (s, 3H), 1.22 (d, J=6.3 Hz, 3H), 1.13(d, J=7.5 Hz, 3H), 0.91 (t, J=7.5 Hz, 9H), 057 (q, J=7.5 Hz, 6H).

PNB-Protected N-Methyl Thioguanidine CP Intermediate 61

Method E; Percent yield: 90%; ¹H NMR (acetone-d₆, 300 MHz): δ 9.60 (brs, 1H), 8.61 (br s, 2H), 8.22 (d, J=8.4 Hz, 2H), 7.79 (d, J=8.4 Hz, 2H),5.52 (d, J=14.1 Hz, 1H), 5.30 (d, J=14.1 Hz, 1H), 4.29 (dd, J=10.5; 3.0Hz, 1H), 4.16-4.08 (m, 1H), 4.03 (d, J=14.7 Hz, 1H), 3.48-3.31 (m, 5H),2.94-2.87 (m, 2H), 2.33 (s, 3H), 1.23 (d, J=7.2 Hz, 3H), 1.19 (d, J=7.2Hz, 3H).

N-Methyl Thioguanidine CP Analog 62

Percent yield: 20%; ¹H NMR (D₂O, 300 MHz): δ 4.20-4.13 (m, 2H), 3.80 (d,J=13.8 Hz, 1H), 3.51 (d, J=13.8 Hz, 1H), 3.40-3.39 (m, 1H), 3.32-3.30(m, 2H), 3.18-3.12 (m, 1H), 3.09-2.92 (m, 2H), 2.47 (s, 3H), 1.22 (d,J=6.9 Hz, 3H), 1.07 (d, J=7.2 Hz, 3H).

Example 20 TES-Protected Prolinol CP Intermediate 58

Method D; Percent yield: 66%; ¹H NMR (CDCl₃, 300 MHz): δ 8.20 (d, J=9.3Hz, 2H), 7.66 (d, J=8.4 Hz, 2H), 5.45 (d, J=14.1 Hz, 1H), 5.22 (d,J=14.1 Hz, 1H), 4.27-4.19 (m, 1H), 4.21 (dd, J=11.4; 3.9 Hz, 1H), 3.96(d, J=15.0 Hz, 1H), 3.65 (dd, J=11.1; 4.2 Hz, 1H), 3.47-3.41 (m, 1H),3.39-3.32 (m, 2H), 3.24 (dd, J=4.8; 2.7 Hz, 1H), 3.00-2.94 (m, 1H),2.70-2.65 (m, 1H), 2.54 (br s, 1H), 2.27-2.18 (m, 1H), 1.98-1.63 (m,4H), 1.25 (d, J=6.0 Hz, 3H), 1.15 (d, J=7.2 Hz, 3H), 0.93 (t, J=7.2 Hz,9H), 0.59 (q, J=7.5 Hz, 6H).

TES-Protected Thioguanidine CP Intermediate 60

Percent yield: 43%; ¹H NMR (CDCl₃, 300 MHz): δ 9.33 (br s, 2H), 8.20 (d,J=9.3 Hz, 2H), 7.67 (d, J=8.7 Hz, 2H), 5.43 (d, J=14.1 Hz, 2H), 5.25 (d,J=14.1 Hz, 2H), 4.30-4.19 (m, 3H), 3.50-3.34 (m, 2H), 3.27-3.25 (m, 1H),3.20-3.07 (m, 3H), 2.95-2.90 (m, 1H), 2.45-2.35 (m, 1H), 2.10-1.98 (m,1H), 1.85-1.70 (m, 3H), 1.21 (d, J=6.9 Hz, 3H), 1.17 (d, J=7.8 Hz, 3H),0.92 (t, J=8.4 Hz, 9H), 0.58 (q, J=8.4 Hz, 6H).

Thioguanidine CP Analog 63

Method F; Percent yield: 15% (from 60); ¹H NMR (D₂O, 400 MHz): δ4.24-4.20 (m, 1H), 4.18 (dd, J=9.6; 2.8 Hz, 1H), 3.83 (d, J=13.2 Hz,1H), 3.46 (d, J=13.2 Hz, 1H), 3.41 (dd, J=6.4; 2.8 Hz, 1H), 3.33-3.26(m, 2H), 3.23-3.16 (m, 1H), 3.10-3.05 (m, 1H), 3.04-2.98 (m, 1H),2.37-2.32 (m, 1H), 2.13-2.08 (m, 1H), 1.86-1.68 (m, 3H), 1.28 (d, J=6.4Hz, 3H), 1.11 (d, J=7.2 Hz, 3H).

Synthesis of the Amine Sidechains Preparation of Sarcosinamide 64

N-Methylglycinamide was prepared from methylamine and α-bromoacetamideas previously described in literature for sarcosinamide hydrochloride,(Marvel, C. S.; Elliott, J. R.; Boettner, F. E.; Yuska, H., J. Am. Chem.Soc. 1946, 68, 1681-1686).

To a stirred solution of bromoacetamide (3.73 g, 27 mmol) in anhydrousTHF (40 mL) at 0° C. under N₂ atmosphere was added a solution ofmethylamine in THF (2.0M in THF, 115 mL, 230 mmol), so as to maintain aninternal temperature below 5° C. The reaction mixture was aged at 0-5°C. for 3 h (monitored by TLC) and then allowed to warm to rt andevaporated under reduced pressure. The crude hydrobromide salt wasdissolved in methanol and the pH adjusted to 8-9 with NaOH solution inmethanol. The solvent was removed under reduced pressure and crudematerial was dried under high vacuum, and then treated with ACN andstirred for several minutes. The insoluble material was filtered off andwashed with ACN (2×10 mL). The combined filtrate was concentrated underreduced pressure, and the residue was purified by flash chromatographyon silica gel (eluent-acetonitrile followed by acetonitrile/water withgradient) to give the desired product 64 in 75% yield.

¹H NMR (DMSO-d₆, 300 MHz): δ 7.28 (br s, 1H), 7.05 (br s, 1H), 3.00 (s,2H), 2.22 (s, 3H).

2-(N-Methylamino)-1-Carbonyl Ethylsulfamide 65

N-Boc-protected sarcosine was activated as its N-hydroxysuccinimidoester and then allowed to react with sulfamide to give, afterdeprotection, the desired product 65.

Step 1

To a stirred solution of N—BOC amino acid (10 mmol) in dry DCM (30 mL)at 0° C. were added N-hydroxysuccinimide (15 mmol) and1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (EDC, 12mmol). The resulting mixture was allowed to warm to room temperature,stirred for ˜20 h (monitoring by TLC) and concentrated under reducedpressure. The residue was then dissolved in ethyl acetate (50 mL),washed with water (4×10 mL) and brine (1 omL), dried over Na₂SO₄,concentrated, and dried under high vacuum to give the hydroxysuccinimidoester in quantitative yield.

¹H NMR (CDCl₃, 300 MHz): δ 4.24 (s, 2H), 3.00 (s, 3H), 2.86 (s, 4H),1.47 (s, 9H).

Step 2

To a solution of the N-Boc-N-hydroxysuccinimido ester of sarcosine (2.86g, 10.0 mmol) in dry DMF (20 mL) was added sulfamide (1.92 g, 20.0 mmol)and the resulting mixture was stirred under nitrogen at 90°-95° C. for 8h (monitoring by TLC). The mixture was then filtered, and the filtratewas evaporated under reduced pressure. The residue was dried under highvacuum and then dissolved in ethyl acetate (20 mL) The resultingsolution was extensively washed with water (monitoring by TLC) and thecombined organics were dried over Na₂SO₄ and concentrated under reducedpressure to give crude material. After drying under high vacuum, thematerial was triturated from ethyl acetate and hexanes and the insolublematerial removed by filtration. The filtrate was evaporated underreduced pressure and the remaining solid was washed with ethyl ether,dissolved in DCM, and the product precipitated by adding hexanes.

Percent yield: 70%; ¹H NMR (CDCl₃, 300 MHz): δ 6.60-6.30 (m, 2H), 3.88(s, 2H), 2.94 (s, 9H).

Step 3

To a stirred solution of Boc-protected amine (1.0 mmol) in DCM (2-5 mL)at 0° C. was added trifluoroacetic acid (95% aqueous solution, 0.5 mL)and the mixture was stirred at 0° C. for 30 minutes, and then allowed towarm to rt and stirred overnight. The volatiles were evaporated underreduced pressure and the residue was treated two times with ethyl etherand evaporated yielding the corresponding amine TFA salt. The salt wasdissolved in methanol and treated with NaOH solution in methanol (pHadjusted to 8-9) and concentrated. The remaining solid was partitionedbetween ethyl acetate and water, the layers separated, and the organiclayer was dried and concentrated to produce the desired product inquantitative yield.

¹H NMR (DMSO-d₆, 300 MHz): δ 7.25 (s, 1H), 7.03 (s, 1H), 3.04 (s, 2H),2.22 (s, 3H).

N-Methyl-N′-PNB Ethylenediamine 66

The synthesis involved the selective protection of the two amine groupsin N-methylethylenediamine, first with ethyltrifluoroacetate and,second, with di-tert-butyl dicarbonate, followed by removal of thetrifluoroacetyl group and reaction of this amine withp-nitrobenzylchloroformate. The synthesis ends with the removal of theBoc-protecting group.

Steps 1 and 2

The procedure used to make N-Boc-N-methyl ethylenediamine was similar tothat previously described in literature (Martins, E. T.; Baruah, H.;Kramarczyk, J.; Saluta, G.; Day, C. S.; Kucera, G. L.; Bierbach, U. J.Med. Chem. 2001, 44, 4492-4496).

Step 3

To a stirred solution of N-Boc-N-methyl ethylenediamine (1.39 g, 8.0mmol) and DIEA (1.05 g, 8.1 mmol) in dry DCM (50 mL) at 0° C. was addeda solution of p-nitrobenzylchloroformate (1.75 g, 8.1 mmol) in dry DCM(10 mL) dropwise over 5-10 minutes. The reaction mixture was stirred atthis temperature for 30 minutes and then allowed to warm to roomtemperature, and stirred overnight (monitored by TLC). The reactionmixture was then washed with 1M aq. NaHCO₃ solution, water, and brine.The organic layer was dried over Na₂SO₄, concentrated, and the remainingresidue was purified by flash chromatography to give the desiredN-Boc-N′-PNB protected intermediate in 73% yield.

¹H NMR (CDCl₃, 300 MHz): δ 8.21 (d, J=8.4 Hz, 2H), 7.50 (d, J=9.0 Hz,2H), 5.19 (s, 2H), 3.38 (br s, 4H), 2.89 (s, 3H), 1.45 (s, 9H).

Step 4

Sidechain 66 was prepared using standard TFA deprotection reactionconditions and purified by flash column chromatography on silica gel.

Percent yield; quantitative; ¹H NMR (acetone-d₆, 300 MHz): δ 8.20 (d,J=8.7 Hz, 2H), 7.62 (d, J=8.1 Hz, 2H), 7.41 (br s, 1H), 5.20 (s, 2H),3.55 (br s, 2H), 3.18 (br s, 2H), 2.74 (s, 3H).

N-Methyl-N′-(Aminosulfonyl)Ethylenediamine 67

This sidechain was prepared by treating N-Boc-N-methylethylenediaminewith an excess of sulfamide in refluxing dioxane followed by N-Bocdeprotection.

Step 1

A solution of N-Boc-N-methylethylenediamine (1 g, 5.74 mmol) andsulfamide (1.1 g, 11.48 mmol) in dioxane (20 mL) was stirred at refluxunder N₂ atmosphere (monitored by TLC). The mixture was then filtered toremove the insoluble material and the filtrate was concentrated underreduced pressure and treated with ethyl acetate. The insoluble materialwas again removed by filtration and the filtrate concentrated underreduced pressure to give the desired product in 96% yield.

¹H NMR (acetone-d₆, 300 MHz): δ 5.91 (br s, 2H), 5.77 (br s, 1H), 3.38(t, J=6.3 Hz, 2H), 3.20 (t, J=6.3 Hz, 2H), 2.85 (s, 3H), 1.40 (s, 9H).

Step 2

Percent yield: 91%; ¹H NMR (acetone-d₆, 300 MHz) δ: 6.62 (br s, 2H),6.34 (br s, 1H), 3.48 (t, J=4.8 Hz, 2H), 3.32 (t, J=4.8 Hz, 2H) 2.80 (s,3H).

1-[2-(N-Methylamino)Ethyl]Imidazole 68

1-[2-(N-methylamino)ethyl]imidazole was synthesized from the mesylate of2-(N-Boc-N-methylamono)ethanol, which was allowed to react withimidazole in the presence of LiHMDS. The starting material was preparedby selective protection of amino group with Boc-anhydride in thepresence of Al₂O₃ according to the procedure described in literature(Yadar, V. K.; Ganesh Babu, K. J. Org. Chem. 2004, 69, 577-580).

Preparation of 1-[2-(N-Boc-N-Methylamino)Ethyl]Imidazole

To a stirred solution of imidazole (1.02 g, 15 mmol) at −30° C. under N₂atmosphere in dry DMF (20 mL) was added LHMDS (1M solution in hexanes,16.0 mL, 16.0 mmol), dropwise, by syringe. The reaction mixture wasstirred at −30° C. for 40 minutes. To this mixture was added a solutionof the mesylate from 2-(N-Boc-N-methylamino)ethanol (freshly preparedfrom 2-(N-Boc-N-methylamino)ethanol, mesyl chloride, and DIEA, (1.75 g,10 mmol)) in DMF (6 mL) via syringe. The resulting mixture was stirredat −30° C. for 30 minutes and then at rt overnight (monitored by TLC).The mixture was then evaporated under reduced pressure and purified byflash chromatography on silica gel to produce the desired Boc-protectedamine in 54% yield.

¹H NMR (CDCl₃, 300 MHz) δ: 7.09 (s, 1H), 6.69 (s, 1H), 6.60 and 6.57(s+s, 1H), 3.76 (t, J=6.0 Hz, 2H), 3.16 (br s, 2H), 2.40 and 2.28 (s+s,3H), 1.10 and 1.05 (s+s, 9H).

Step 2: Preparation of Imidazole 68

Percent yield: 83%; ¹H NMR (CDCl₃, 300 MHz) δ: 7.51 (s, 1H), 7.07 (s,1H), 6.96 (s, 1H), 4.07 (t, J=6.0 Hz, 2H), 2.93 (t, J=6.0 Hz, 2H), 2.43(s, 3H), 1.13 (br s, 1H).

N-Methylaminoethyl-N′,N″-bis-PNB-Oxycarbonyl-Guanidine 69

This sidechain was prepared by displacement of pyrazole from thebis-PNB-protected 1H-pyrazole-carboxamidine (shown above), followed byremoval of the Boc protecting group.

Step 1: N-Boc-N-Methylaminoethyl-N′,N″-Bis-PNB Guanidine

To a solution of N-Boc-N-methylethylenediamine (627 mg, 3.6 mmol) in dryDMF (10 mL) was added bis-PNB-protected pyrazole-1-carboxamidine (750mg, 1.6 mmol) and the resulting mixture was stirred at rt for 3 days(monitoring by TLC). The solvent was then removed under reduced pressureand purified by flash chromatography to produce the desiredBoc-protected amine in 50% yield.

¹H NMR (CDCl₃, 300 MHz) δ: 11.70 (s.1H), 8.47 and 8.35 (s+s, 1H), 8.14(d, J=8.1 Hz, 2H), 8.11 (d, J=8.1 Hz, 2H), 7.48 (d, J=8.1 Hz, 2H), 7.46(d, J=8.1 Hz, 2H), 5.21 (s, 2H), 5.15 (s, 2H), 3.56-3.51 (m, 2H), 3.41(t, J=5.4 Hz, 2H), 2.81 (s, 3H), 1.34 (s, 9H).

Step 2: Amine 69

Percent yield: 89%; ¹H NMR (CDCl₃, 300 MHz) δ: 11.63 (s, 1H), 9.80 (s,1H), 8.65 (t, J=6.0 Hz, 1H), 8.19 (d, J=4.5 Hz, 2H), 8.16 (d, J=4.5 Hz,2H), 7.51 (d, J=5.7 Hz, 2H), 7.48 (d, J=5.1 Hz, 2H), 5.23 (s, 2H), 5.20(s, 2H), 3.76 (br s, 2H), 2.70 (s, 3H)

2-(N-Methylaminomethyl)Thiazole 70

To a flask containing dry DCM (60 mL) and 4 A molecular sieves wereadded methylamine (2.0M solution in THF, 4.0 mL, 8.0 mmol), Na₂SO₄ (1.42g, 10.0 mmol), and 2-thiazolecarboxaldehyde (0.452 g, 4.0 mmol). Thereaction mixture was stirred at room temperature and monitored by ¹HNMR. Upon completion, the reaction mixture filtered to remove the Na₂SO₄and molecular sieves and the filtrate was concentrated under reducedpressure. The crude imine was dissolved in absolute EtOH (40 mL),treated with NaBH₄ (0.227 g, 6.0 mmol) and aged for several hours at rt(monitored by TLC). The reaction mixture was then quenched with water (8mL) and concentrated under reduced pressure. The residue was treatedwith brine, extracted with DCM (3×50 mL), and the combined organiclayers were dried, and evaporated under reduced pressure to give thedesired product in 69% yield.

¹H NMR (CDCl₃, 300 MHz) δ: 7.69 (d, J=3.6 Hz, 1H), 7.24 (d, J=3.6 Hz,1H0, 4.06 (s, 2H), 2.49 (s, 3H), 1.80 (br s, 1H).

2-[(Aminosulfonyl)Aminomethyl]piperazine 71

The synthesis involved preparation of1,4-dibenzyl-piperazine-2-carboxylic acid amide, reduction of the amideto the amine with LAH, conversion of the amine to the sulfonamide, anddeprotection of the bis-benzyl protecting groups.

Step 1: 1,4-Dibenzy piperazine-2-Carboxylic Acid Amide

This compound was prepared in 75% yield according to the proceduredescribed in literature (Butts, C. P.; Jazdzyk, M. Chem. Commu. 2003,(13) 1530-1531)

Step 2: 1,4-Dibenzyl-2-aminomethyl-piperazine

To a stirred suspension of LAH (0.9 g, 25.9 mmol) in dry THF (150 mL)was added 1,4-dibenzyl-piperazine-2-carboxylic acid amide (4.0 g, 12.93mmol) and the mixture was stirred under reflux for 20 h (monitored byTLC). The stirred mixture was cooled with an ice-bath and quenched withRochelle's salt (6 mL), warmed to room temperature, and the suspensionwas poured over a pad of silica gel (20 mm). The celite was washed with15% MeOH in CH₂Cl₂ with 1% NH₄OH (3×50 mL) and eluted through pad ofsilica gel. The eluent was concentrated under reduced pressure to givethe desired product in 99% yield.

¹H NMR (CDCl₃, 300 MHz): δ 7.32-7.31 (m, 10H), 4.05 (d, J=13.8 Hz, 1H),3.52 and 3.47 (AB_(q), J=12.9 Hz, 2H), 3.27 (d, J=13.8 Hz, 1H), 3.06(dd, J=13.2; 5.7 Hz, 1H), 2.82-2.69 (m, 3H), 2.63-2.58 (m, 1H),2.50-2.43 (m, 1H), 2.36-2.16 (m, 3H).

Step 3: 1,4-Dibenzyl-2-[(Aminosulfonyl)Aminomethyl]piperazine

To a solution of 1,4-dibenzyl-2-aminomethyl-piperazine (2.5 g, 8.46mmol) in dioxane (50 mL), was added sulfamide (1.63 g, 16.9 mmol) andthe resulting mixture was stirred at 95° C. under N₂ atmosphere forseveral hours (monitored by TLC). The solvent was removed under reducedpressure and the crude material was purified by flash chromatography onsilica gel using acetonitrile/water with gradient as eluent.

Percent yield: 58%; ¹H NMR (CDCl₃, 300 MHz) δ: 7.37-7.29 (m, 10H), 5.64(br s, 1H), 4.24 (br s, 2H), 3.94 (d, J=12.9 Hz, 1H), 3.59-3.46 (m, 4H),3.13 (d, J=12.9 Hz, 1H), 2.96-2.91 (m, 1H), 2.81-2.76 (m, 1H).

Step 4: 2-[(Aminosulfonyl)Aminomethyl]piperazine

A mixture of 1,4-dibenzyl-piperazine-2-methylsulfamide (1.82 g, 4.87mmol) in MeOH (20 mL) and 10% Pd/C (180 mg) was treated with hydrogengas (65 psi) for 27 h at rt. The mixture was then filtered through a padof Celite and the celite washed well with methanol. The filtrate wasconcentrated under reduced pressure to give the desired product 71 in96% yield.

¹H NMR (DMSO-d₆, 300 MHz) δ: 6.48 (br s, 1H), 6.42 (br s, 1H), 3.20-3.13(m, 2H), 2.77-2.62 (m, 3H), 2.57-2.50 (m, 2H), 2.47-2.39 (m with DMSO-d₆peak, 1H), 2.11 (dd, J=11.4; 9.6 Hz, 1H).

(2S)-2-[(Aminosulfonyl)aminocarbonyl]pyrrolidine 72

(2S)-1-Cbz-2-[(Aminosulfonyl)Aminocarbonyl]Pyrrolidine

Percent yield: 42%; ¹H NMR (CDCl₃, 300 MHz): δ 7.34-7.26 (m, 5H), 6.76(s, 1H), 6.42-6.26 (m, 2H), 5.11 (s, 2H), 4.31 (br s, 1H), 3.52-3.40 (m,2H), 2.20-2.11 (m, 2H), 1.95-1.86 (m, 3H).

(2S)-2-[(Aminosulfonyl)Aminocarbonyl]Pyrrolidine 72

Percent yield: 95%; ¹H NMR (CDCl₃, 300 MHz) δ: 7.40 (br s, 1H), 6.16 (brs, 1H), 3.69 (dd, J=9.0; 5.1 Hz), 3.06-2.85 (m, 2H), 2.35 (br s, 1H),2.17-2.05 (m, 1H), 1.94-1.83 (m, 1H), 1.80-1.59 (m, 2H).

Synthesis of Secondary Amines by Alkylation

The synthesis of a series of 2-, 3- or 4-(alkylamino)alkylpyridines wasachieved by protection of 2-, 3- or 4-aminoalkylpyridines with Bocanhydride in t-butyl alcohol followed by treatment with sodium hydridein DMF and alkylation with various alkyl halides. Lastly, removal of theBoc protecting group was accomplished with 95% TFA in DCM. Thissynthetic strategy was first reported in literature for preparation of2-(alkylamino)pyridines (Krein, D. M.; Lowary, T. L. J. Org. Chem. 2002,67, 4965-4967).

Examples of Prepared Amines:

2-(Methylaminomethyl)-5-methylpyrazine—66% overall yield; ¹H NMR (CDCl₃,300 MHz) δ: 8.42 (s, 1H), 8.35 (s, 1H), 3.83 (s, 2H), 2.50 (s, 3H), 2.43(s, 3H).

2-(methylaminomethyl)pyridine—55% overall yield; ¹H NMR (CDCl₃, 300 MHz)δ: 8.58 (s, 1H), 7.71 (t, J=8.4 Hz, 1H), 7.31-7.24 (m, 2H), 4.01 (s,2H), 2.54 (s, 3H).

3-[(4-thiazolomethylamino)methyl]pyridine—42% overall yield; ¹H NMR(CDCl₃, 300 MHz) δ: 8.79 (d, J=1.2 Hz, 1H), 8.57 (d, J=4.2 Hz, 1H), 8.08(dt, J=8.1; 2.4 Hz, 7.35 (d, J=6.9 Hz, 1H), 7.24 (br s, 1H), 7.19-7.15(m, 1H), 4.06 (s, 2H), 3.99 (s, 2H), 2.80 (s, 1H).

Dilution Antimicrobial Susceptibility Tests

The agar dilution method for determining the antimicrobialsusceptibility was carried out using an agar dilution method withMueller-Hinton agar (see, M7-A5, Vol. (2), 2000). A final inoculum of104 CFU was applied with a replicating device. Broth dilution tests wereperformed with 5×105 CFU in tubes containing 1 mL of broth. Incubationof test tubes containing agar and broth was done at 35 C for 18 h. Thesusceptibilities of streptococci were determined by Mueller-Hinton agarsupplemented with 5% sheep blood, and the susceptibility of anaerobicspecies was determined with brucella agar supplemented with 5% sheepblood, hemin, and vitamin K. Incubation of anaerobic cultures was donefor 48 h in jars. The susceptibilities of methicillin-resistantstaphylococci were determined on Mueller-Hinton agar or in brothsupplemented with 3% NaCl. All assays were run with the indicatedcontrol strains, available from the American Type Culture Collection,Rockville, Md.). Results of the antimicrobial susceptibility tests ofcompounds 9-63 against Gram-negative organisms are shown in Table I.

The compositions, methods and/or processes disclosed and claimed hereincan be made and executed without undue experimentation in light of thepresent disclosure. While the compositions and methods of this inventionhave been described in terms of preferred embodiments, it will beapparent to those of skill in the art that variations may be applied tothe compositions, methods and/or processes and in the steps or in thesequence of steps of the methods described herein without departing fromthe concept and scope of the invention. More specifically, it will beapparent that certain agents which are both chemically andphysiologically related may be substituted for the agents describedherein while the same or similar results would be achieved. All suchsimilar substitutes and modifications apparent to those skilled in theart are deemed to be within the scope and concept of the invention.

1-39. (canceled)
 40. A compound selected from the group consisting of:

or a pharmaceutically acceptable salt of prodrug thereof.
 41. Thecompound of claim 40, wherein the compound is:

or a pharmaceutically acceptable salt of prodrug thereof.
 42. Thecompound of claim 40, wherein the compound is:

or a pharmaceutically acceptable salt of prodrug thereof.
 43. Thecompound of claim 40, wherein the compound is:

or a pharmaceutically acceptable salt of prodrug thereof.
 44. Thecompound of claim 40, wherein the compound is:

or a pharmaceutically acceptable salt of prodrug thereof.
 45. Thecompound of claim 40, wherein the compound is:

or a pharmaceutically acceptable salt of prodrug thereof.
 46. Thecompound of claim 40, wherein the compound is:

or a pharmaceutically acceptable salt of prodrug thereof.
 47. Thecompound of claim 40, wherein the compound is:

or a pharmaceutically acceptable salt of prodrug thereof.
 48. Thecompound of claim 40, wherein the compound is:

or a pharmaceutically acceptable salt of prodrug thereof.
 49. Thecompound of claim 40, wherein the compound is:

or a pharmaceutically acceptable salt of prodrug thereof.
 50. Apharmaceutical composition comprising a compound of claim 40 and apharmaceutically acceptable carrier.
 51. A method of preventing ortreating an infection by a gram negative bacteria in a host comprisingadministering to the host a therapeutic amount of a compound of claim40, optionally in a pharmaceutically acceptable carrier.
 52. The methodof claim 51, wherein the host is a human.
 53. The method of claim 51,wherein the infection is by a drug resistant bacterial strain.
 54. Themethod of claim 51, wherein the infection is by a multiple-drugresistant strain.
 55. The method of claim 51, wherein the compound isadministered in combination or alternation with at least one otherantimicrobial agent.
 56. The method of claim 51, wherein the compound isadministered in combination with a β-lactamase inhibiting agent.
 57. Aprocess for preparing a β-methyl carbapenem analogs comprising: (a)preparing or obtaining a carbapenem intermediate of the structure (A)

wherein P is H, hydroxy, halogen, or hydroxyl protected by a hydroxylprotecting group; R is H or alkyl; R¹ is H or alkyl; P′ is a suitablecarboxyl protecting group; and R′ is an alkyl or substituted alkyl; (b)coupling the compound of structure (A) with a moiety with a freehydroxyl or a mono- or di-substituted amine to obtain a β-methylcarbapenem analog; and (c) optionally deprotecting the β-methylcarbapenem analog.
 58. The process of claim 57, wherein the compound ofstructure (A) is


59. The process of claim 57, wherein the moiety with a free hydroxyl isan aromatic alcohol or a heteroaromatic alcohol.
 60. The process ofclaim 57, wherein the mono- or di-substituted amine is an aromatic amineor a heteroaromatic amine.